Methods for making compositions for improving skin conditions

ABSTRACT

The present specification discloses fluid compositions comprising a matrix polymer and stabilizing component, methods of making such fluid compositions, and methods of treating skin conditions in an individual using such fluid compositions.

This patent application is a continuation of U.S. patent applicationSer. No. 14/078,057, filed Nov. 12, 2013, which is a divisional of U.S.patent application Ser. No. 12/777,106, now U.S. Pat. No. 8,586,562,issued Nov. 19, 2013, which claims priority pursuant to 35 U.S.C.§119(e) to U.S. Provisional Patent Application Ser. No. 61/313,664 filedMar. 12, 2010, the entire disclosure of each of these applications beingincorporated herein by this reference.

Dermal fillers are useful in soft tissue and dermal correction. Onecommon polymer used in dermal filler compositions is hyaluronan, alsoknown as hyaluronic acid (HA). Although exhibiting excellentbiocompatibility and affinity for water molecules, in its natural state,hyaluronan exhibits poor biomechanical properties as a dermal filler.Tezel and Fredrickson, The Science of Hyaluronic Acid Dermal Fillers, JCosmet Laser Ther. 10(1): 35-42 (2008); Kablik, et al., ComparativePhysical Properties of Hyaluronic Acid Dermal Fillers, Dermatol Surg. 35Suppl 1: 302-312 (2009); Beasley, et al., Hyaluronic Acid Fillers: AComprehensive Review, Facial Plast Surg. 25(2): 86-94 (2009); each ofwhich is hereby incorporated by reference in its entirety. One primaryreason is that this polymer is soluble and is cleared rapidly whenadministered into a skin region. Tezel, supra, 2008; Kablik, supra,2009; Beasley, supra, 2009. This in vivo clearance is primarily achievedby degradation, principally enzymatic degradation via hyaluronidase andchemical degradation via free-radicals. To minimize the effect of thesein vivo degradation pathways, matrix polymers like hyaluronan arecrosslinked to one another to form a hydrogel. Because hydrogels aremore solid substance that are readily soluble, dermal fillers comprisingsuch crosslinked matrix polymers remain in place at the implant site.Tezel, supra, 2008; Kablik, supra, 2009; Beasley, supra, 2009. Acrosslinked matrix polymer like hyaluronan is also more suitable as acomponent of a dermal filler because it's more solid nature improves themechanical properties of the filler, allowing the filler to better liftand fill a skin region. Tezel, supra, 2008; Kablik, supra, 2009;Beasley, supra, 2009.

Hyaluronan is abundant in the different layers of the skin, where it hasmultiple functions such as to ensure good hydration, assist in theorganization of the collagen matrix, and act as a filler materialassisting the organization of the extracellular matrix. However, withage, the quantity of hyaluronan present in the skin decreases. This lossof hyaluronan results in various skin conditions such as, e.g., skindehydration, lack of skin elasticity, skin roughness, lack of skintautness, skin stretch line and/or marks, skin paleness, skin wrinkles,and the like. As such, it would be desirable to have a skin therapy thatcan replace the endogenous matrix polymers that are lost with age inorder to treat these skin conditions. However, current dermal fillerscomprising hydrogels of crosslinked matrix polymers like crosslinkedhyaluronan cannot be used to replace the lost endogenous polymersbecause the crosslinking prevents the ability of these polymers tointegrate into the extracellular matrix. However, as discussed above,although uncrosslinked matrix polymers like hyaluronan are soluble, andas such, could integrate into the extracellular matrix and replace lostendogenous hyaluronan, uncrosslinked matrix polymers are rapidly clearedfrom the body by in vivo degradation pathways. Thus, what is needed is afluid composition comprising uncrosslinked matrix polymers that includean additional stabilizing component that reduces or prevents matrixpolymer degradation.

The fluid compositions disclosed in the present specification achievethis goal. Such fluid compositions comprise a matrix polymer and astabilizing component that reduces or prevents in vivo degradation ofthe matrix polymer. Administration of the disclosed fluid compositionsimproves skin conditions such as, e.g., hydration and the cutaneouselasticity by compensating for the loss of the endogenous polymer.

Thus, aspects of the present specification provide a fluid compositioncomprising a matrix polymer and a stabilizing component. Matrix polymersuseful to make such fluid compositions include, without limitation, aglycosaminoglycan (like chondroitin sulfate, dermatan sulfate, keratansulfate, hyaluronan) and lubricin. Stabilizing components useful to makesuch fluid compositions include, without limitation, polyols andflavonoids.

Other aspects of the present specification provide a method of making afluid composition disclosed in the present specification. In an aspect,a method for making a fluid composition comprises the steps of: a)combining a stabilizing component with a physiologically-acceptablebuffer to make a stabilizing component-buffered solution; and b)combining a matrix polymer with the stabilizing component-bufferedsolution to hydrate the matrix polymer. In another aspect, a method formaking a fluid composition comprises the steps of: a) combining astabilizing component with a physiologically-acceptable buffer to make astabilizing component-buffered solution; b) combining a matrix polymerwith the stabilizing component-buffered solution to hydrate the matrixpolymer, and; c) sizing the fluid composition. This method may, or maynot, further comprise a step comprising titrating a stabilizingcomponent-buffered solution to obtain a desired pH after step (a); astep comprising filtering the stabilizing component-buffered solutionafter step (a); a step (b) where combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymeroccurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time; a step (b) where combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymeroccurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time and then followed by a rest for a relative long period of time;a step (b) where combining a matrix polymer with the stabilizingcomponent-buffered solution to hydrate the matrix polymer occurs bymixing the matrix polymer with the stabilizing component-bufferedsolution at a low speed for a relatively long period of time and then bymixing the matrix polymer with the stabilizing component-bufferedsolution using a cycle of alternating periods of agitation for arelatively short period of time followed by periods of rest for arelatively long period of time; a step (b) where combining a matrixpolymer with the stabilizing component-buffered solution to hydrate thematrix polymer occurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time and then by mixing the matrix polymer with the stabilizingcomponent-buffered solution using a cycle of alternating periods ofagitation for a relatively short period of time followed by periods ofrest for a relatively long period of time, and then followed by a restfor a relative long period of time; a step comprising degassing a fluidcomposition after step (b) or step (c); a step comprising filling asyringe with a fluid composition after step (c); and/or a stepcomprising sterilizing a syringe filled with a fluid composition afterstep (c).

Yet other aspects of the present specification provide a fluidcomposition disclosed in the present specification made by a methoddisclosed in the present specification.

Still other aspects of the present specification provide a method ofimproving a condition of skin in an individual in need thereof, themethod comprising the steps of administering a fluid compositiondisclosed in the present specification into a dermal region of theindividual, wherein the administration improves the condition. Skinconditions treated by the disclosed fluid compositions include, withoutlimitation, skin dehydration, a lack of skin elasticity, skin roughness,a lack of skin tautness, a skin stretch line or mark, skin paleness,and/or skin wrinkles.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B are graphs showing the results of hyaluronan polymerdegradation with and without mannitol. FIG. 1A shows a graph plottingG′G″ over time. FIG. 1B shows a graph plotting dynamic viscosity overtime. SGM181a is a fluid composition comprising 13.5 mg/mL hyaluronanpolymer without mannitol (control) and SGM181b is a fluid compositioncomprising 13.5 mg/mL hyaluronan polymer and 5% mannitol.

FIG. 2 is a graph showing the effects of increasing stabilizingconcentration on the dynamic viscosity of a fluid composition comprisinga matrix polymer. The graph plots dynamic viscosity (Pa·s) over time(s). SGM173a is a fluid composition comprising 13.5 mg/mL hyaluronanpolymer without mannitol (control), SGM173c is a fluid compositioncomprising 13.5 mg/mL hyaluronan polymer and 5% mannitol and SGM173d isa fluid composition comprising 13.5 mg/mL hyaluronan polymer and 9%mannitol.

DETAILED DESCRIPTION

Aspects of the present specification provide, in part, a fluidcomposition comprising a matrix polymer and a stabilizing component. Asused herein, the term “fluid” refers to a continuous, amorphoussubstance whose molecules move freely past one another. A fluid cannotsustain a shearing force when at rest and undergoes a continuous changein shape when subjected to such a force. It should be noted, thatalthough the compositions disclosed in the present specification arefluid in nature due to the presence of uncross linked matrix polymers,such fluid compositions, may, although may not, include cross linkedmatrix polymers, which, by its nature, is a gel or other solidsubstance. As such, certain fluid compositions disclosed in the presentspecification exhibit viscoelastic properties.

Aspects of the present specification provide, in part, a fluidcomposition comprising a matrix polymer. As used herein, the tem “matrixpolymer” refers to a polymer that can become part of or function as anextracellular matrix polymer and pharmaceutically acceptable saltsthereof. Non-limiting examples of a matrix polymer include aglycosaminoglycan like chondroitin sulfate, dermatan sulfate, keratansulfate, hyaluronan; lubricants; and collagens. Non-limiting examples ofa pharmaceutically acceptable salt of a matrix polymer includes sodiumsalts, potassium salts, magnesium salts, calcium salts, and combinationsthereof.

Aspects of the present specification provide, in part, a fluidcomposition comprising a glycosaminoglycan. As used herein, the term“glycosaminoglycan” is synonymous with “GAG” and “mucopolysaccharide”and refers to long unbranched polysaccharides consisting of a repeatingdisaccharide units. The repeating unit consists of a hexose (six-carbonsugar) or a hexuronic acid, linked to a hexosamine (six-carbon sugarcontaining nitrogen) and pharmaceutically acceptable salts thereof.Members of the GAG family vary in the type of hexosamine, hexose orhexuronic acid unit they contain, such as, e.g., glucuronic acid,iduronic acid, galactose, galactosamine, glucosamine) and may also varyin the geometry of the glycosidic linkage. Any glycosaminoglycan isuseful in the compositions disclosed in the present specification withthe proviso that the glycosaminoglycan improves a condition of the skin,such as, e.g., hydration or elasticity. Table 1 lists representativeGAGs.

TABLE 1 Examples of GAGs Glycosidic Hexuronic linkage Name acid/HexoseHexosamine geometry Unique features Chondroitin GlcUA or GalNAc or-4GlcUAβ1- Most prevalent GAG sulfate GlcUA(2S) GalNAc(4S) or 3GalNAcβ1-GalNAc(6S) or GalNAc(4S, 6S) Dermatan GlcUA or GalNAc or -4IdoUAβ1-Distinguished from sulfate IdoUA or GalNAc(4S) or 3GalNAcβ1- chondroitinsulfate by the IdoUA(2S) GalNAc(6S) or presence of iduronic acid,GalNAc(4S, 6S) although some hexuronic acid monosaccharides may beglucuronic acid. Keratan Gal or GlcNAc or -3Gal(6S)β1- Keratan sulfatetype II may be sulfate Gal(6S) GlcNAc(6S) 4GlcNAc(6S)β1- fucosylated.Heparin GlcUA or GlcNAc or -4IdoUA(2S)α1- Highest negative chargeIdoUA(2S) GlcNS or 4GlcNS(6S)α1- density of any known GlcNAc(6S) orbiological molecule GlcNS(6S) Heparan GlcUA or GlcNAc or -4GlcUAβ1-Highly similar in structure to sulfate IdoUA or GlcNS or 4GlcNAcα1-heparin, however heparan IdoUA(2S) GlcNAc(6S) or sulfates disaccharideunits GlcNS(6S) are organised into distinct sulfated and non-sulfateddomains. Hyaluronan GlcUA GlcNAc -4GlcUAβ1- The only GAG that is3GlcNAcβ1- exclusively non-sulfated GlcUA = β-D-glucuronic acidGlcUA(2S) = 2-O-sulfo-β-D-glucuronic acid IdoUA = α-L-iduronic acidIdoUA(2S) = 2-O-sulfo-α-L-iduronic acid Gal = β-D-galactose Gal(6S) =6-O-sulfo-β-D-galactose GalNAc = β-D-N-acetylgalactosamine GalNAc(4S) =β-D-N-acetylgalactosamine-4-O-sulfate GalNAc(6S) =β-D-N-acetylgalactosamine-6-O-sulfate GalNAc(4S, 6S) =β-D-N-acetylgalactosamine-4-O, 6-O-sulfate GlcNAc =α-D-N-acetylglucosamine GlcNS = α-D-N-sulfoglucosamine GlcNS(6S) =a-D-N-sulfoglucosamine-6-O-sulfate

Aspects of the present specification provide, in part, a fluidcomposition comprising a chondroitin sulfate. As used herein, the term“chondroitin sulfate” refers to an unbranched, sulfated GAG of variablelength comprising disaccharides of two alternating monosaccharides ofD-glucuronic acid (GlcA) and N-acetyl-D-galactosamine (GalNAc) andpharmaceutically acceptable salts thereof. A chondroitin sulfate mayalso include D-glucuronic acid residues that are epimerized intoL-iduronic acid (IdoA), in which case the resulting disaccharide isreferred to as dermatan sulfate. A chondroitin sulfate polymer can havea chain of over 100 individual sugars, each of which can be sulfated invariable positions and quantities. Chondroitin sulfate is an importantstructural component of cartilage and provides much of its resistance tocompression. Any chondroitin sulfate is useful in the compositionsdisclosed in the present specification with the proviso that thechondroitin sulfate improves a condition of the skin, such as, e.g.,hydration or elasticity. Non-limiting examples of pharmaceuticallyacceptable salts of chondroitin sulfate include sodium chondroitinsulfate, potassium chondroitin sulfate, magnesium chondroitin sulfate,calcium chondroitin sulfate, and combinations thereof.

Aspects of the present specification provide, in part, a fluidcomposition comprising a keratan sulfate. As used herein, the term“keratan sulfate” refers to a GAG of variable length comprisingdisaccharide units, which themselves include β-D-galactose andN-acetyl-D-galactosamine (GalNAc) and pharmaceutically acceptable saltsthereof. Disaccharides within the repeating region of keratan sulfatemay be fucosylated and N-Acetylneuraminic acid caps the end of thechains. Any keratan sulfate is useful in the compositions disclosed inthe present specification with the proviso that the keratan sulfateimproves a condition of the skin, such as, e.g., hydration orelasticity. Non-limiting examples of pharmaceutically acceptable saltsof keratan sulfate include sodium keratan sulfate, potassium keratansulfate, magnesium keratan sulfate, calcium keratan sulfate, andcombinations thereof.

Aspects of the present specification provide, in part, a fluidcomposition comprising a hyaluronan. As used herein, the term“hyaluronic acid” is synonymous with “HA”, “hyaluronic acid”, and“hyaluronate” refers to an anionic, non-sulfated glycosaminoglycanpolymer comprising disaccharide units, which themselves includeD-glucuronic acid and D-N-acetylglucosamine monomers, linked togethervia alternating β-1,4 and β-1,3 glycosidic bonds and pharmaceuticallyacceptable salts thereof. Hyaluronan can be purified from animal andnon-animal sources. Polymers of hyaluronan can range in size from about5,000 Da to about 20,000,000 Da. Any hyaluronan is useful in thecompositions disclosed in the present specification with the provisothat the hyaluronan improves a condition of the skin, such as, e.g.,hydration or elasticity. Non-limiting examples of pharmaceuticallyacceptable salts of hyaluronan include sodium hyaluronan, potassiumhyaluronan, magnesium hyaluronan, calcium hyaluronan, and combinationsthereof.

Aspects of the present specification provide, in part, a fluidcomposition comprising a lubricin. As used herein, the term “lubricin”refers to a large, water soluble glycoprotein encoded by the PRG4 geneand pharmaceutically acceptable salts thereof. It has a molecular weightof 206,000 Da and comprises approximately equal proportions of proteinand glycosaminoglycans. The structure of lubricin molecule is that of apartially extended flexible rod and, in solution, occupies a smallerspatial domain than would be expected from structural predictions. Thischaracteristic may aid in the molecule's boundary lubricating ability.Lubricin is present in synovial fluid and on the surface (superficiallayer) of articular cartilage and therefore plays an important role injoint lubrication and synovial homeostasis. Any lubricin is useful inthe compositions disclosed in the present specification with the provisothat the lubricin improves a condition of the skin, such as, e.g.,hydration or elasticity. Non-limiting examples of pharmaceuticallyacceptable salts of lubricin include sodium lubricin, potassiumlubricin, magnesium lubricin, calcium lubricin, and combinationsthereof.

Aspects of the present specification provide, in part, a fluidcomposition comprising a crosslinked matrix polymer. As sued herein, theterm “crosslinked” refers to the intermolecular bonds joining theindividual polymer molecules, or monomer chains, into a more stablestructure like a gel. As such, a crosslinked matrix polymer has at leastone intermolecular bond joining at least one individual polymer moleculeto another one. Matrix polymers disclosed in the present specificationmay be crosslinked using dialdehydes and disulfides crosslinking agentsincluding, without limitation, divinyl sulfones, diglycidyl ethers, andbis-epoxides. Non-limiting examples of hyaluronan crosslinking agentsinclude divinyl sulfone (DVS), 1,4-butanediol diglycidyl ether (BDDE),1,2-bis(2,3-epoxypropoxy)ethylene (EGDGE), 1,2,7,8-diepoxyoctane (DEO),biscarbodiimide (BCD), adipic dihydrazide (ADH),bis(sulfosuccinimidyl)suberate (BS), hexamethylenediamine (HMDA),1-(2,3-epoxypropyl)-2,3-epoxycyclohexane, or combinations thereof.

Aspects of the present specification provide, in part, a fluidcomposition comprising a crosslinked matrix polymer having a degree ofcrosslinking. As used herein, the term “degree of crosslinking” refersto the percentage of matrix polymer monomeric units that are bound to across-linking agent, such as, e.g., the disaccharide monomer units ofhyaluronan. Thus, a fluid composition that that has a crosslinked matrixpolymer with a 4% degree of crosslinking means that on average there arefour crosslinking molecules for every 100 monomeric units. Every otherparameter being equal, the greater the degree of crosslinking, theharder the gel becomes. Non-limiting examples of a degree ofcrosslinking include about 1% to about 15%.

Aspects of the present specification provide, in part, a fluidcomposition comprising a uncrosslinked matrix polymer. As used herein,the term “uncrosslinked” refers to a lack of intermolecular bondsjoining the individual matrix polymer molecules, or monomer chains. Assuch, an uncrosslinked matrix polymer is not linked to any other matrixpolymer by an intermolecular bond.

Aspects of the present specification provide, in part, a fluidcomposition comprising a substantially uncrosslinked matrix polymer. Assued herein, the term “substantially uncrosslinked” refers to thepresence of uncrosslinked matrix polymers in a fluid compositiondisclosed in the present specification at a level of at least 90% byweight of the composition, with the remaining at most 10% by weight ofthe composition being comprised of other components includingcrosslinked matrix polymers. The matrix polymer included in a fluidcomposition disclosed in the present specification exhibit a low degreeof cross-linking in order to remain water soluble.

Aspects of the present specification provide, in part, a fluidcomposition that is essentially free of a crosslinked matrix polymer. Asused herein, the term “essentially free” (or “consisting essentiallyof”) refers to a fluid composition where only trace amounts ofcross-linked matrix polymers can be detected.

Aspects of the present specification provide, in part, a fluidcomposition that is entirely free of a crosslinked matrix polymer. Asused herein, the term “entirely free” refers to a fluid composition thatwithin the detection range of the instrument or process being used,crosslinked matrix polymers cannot be detected or its presence cannot beconfirmed.

Aspects of the present specification provide, in part, a fluidcomposition comprising a ratio of crosslinked matrix polymer anduncrosslinked polymer. This ratio of crosslinked and uncrosslinkedmatrix polymer is also known as the gel:fluid ratio. Any gel:fluid ratiois useful in making the fluid compositions disclosed in the presentspecification with the proviso that such ratio produces a fluidcomposition disclosed in the present specification that improves a skincondition as disclosed in the present specification. Non-limitingexamples of gel:fluid ratios include 100:0, 98:2, 90:10, 75:25, 70:30,60:40, 50:50, 40:60, 30:70, 25:75, 10:90; 2:98, and 0:100.

Aspects of the present specification provide, in part, a fluidcomposition comprising a matrix polymer having a mean molecular weight.As used herein, the term “molecular weight” refers to the sum of theatomic weights of the atoms in a molecule. For example, that of methane(CH₄) is 16.043 g/mol, the atomic weights being carbon=12.011 g/mol,hydrogen=1.008 g/mol.

Thus, in an embodiment, a fluid composition comprises a substantiallyuncrosslinked matrix polymer. In other aspects of this embodiment, afluid composition comprises an uncrosslinked matrix polymer where theuncrosslinked matrix polymer represents, e.g., about 90% by weight,about 91% by weight, about 92% by weight, about 93% by weight, about 94%by weight, about 95% by weight, about 96% by weight, about 97% byweight, about 98% by weight, or about 99%, or about 100% by weight, ofthe total matrix polymer present in the composition. In yet otheraspects of this embodiment, a fluid composition comprises anuncrosslinked matrix polymer where the uncrosslinked matrix polymerrepresents, e.g., at least 90% by weight, at least 91% by weight, atleast 92% by weight, at least 93% by weight, at least 94% by weight, atleast 95% by weight, at least 96% by weight, at least 97% by weight, atleast 98% by weight, or at least 99% by weight, of the total matrixpolymer present in the composition. In still other aspects of thisembodiment, a fluid composition comprises an uncrosslinked matrixpolymer where the uncrosslinked matrix polymer represents, e.g., about90% to about 100% by weight, about 93% to about 100% by weight, about95% to about 100% by weight, or about 97% to about 100% by weight, ofthe total matrix polymer present in the composition.

In another embodiment, a fluid composition comprises a substantiallyuncrosslinked glycosaminoglycan. In aspects of this embodiment, a fluidcomposition comprises a substantially uncrosslinked chondroitin sulfatepolymer, a substantially uncrosslinked chondroitin sulfate polymer, or asubstantially uncrosslinked hyaluronan polymer. In other aspects of thisembodiment, a fluid composition comprises an uncrosslinkedglycosaminoglycan where the uncrosslinked glycosaminoglycan represents,e.g., about 90% by weight, about 91% by weight, about 92% by weight,about 93% by weight, about 94% by weight, about 95% by weight, about 96%by weight, about 97% by weight, about 98% by weight, or about 99%, orabout 100% by weight, of the total glycosaminoglycan present in thecomposition. In yet other aspects of this embodiment, a fluidcomposition comprises an uncrosslinked glycosaminoglycan where theuncrosslinked glycosaminoglycan represents, e.g., at least 90% byweight, at least 91% by weight, at least 92% by weight, at least 93% byweight, at least 94% by weight, at least 95% by weight, at least 96% byweight, at least 97% by weight, at least 98% by weight, or at least 99%by weight, of the total glycosaminoglycan present in the composition. Instill other aspects of this embodiment, a fluid composition comprises anuncrosslinked glycosaminoglycan where the uncrosslinkedglycosaminoglycan represents, e.g., about 90% to about 100% by weight,about 93% to about 100% by weight, about 95% to about 100% by weight, orabout 97% to about 100% by weight, of the total glycosaminoglycanpresent in the composition.

In yet another embodiment, a fluid composition comprises a substantiallyuncrosslinked lubricin. In aspects of this embodiment, a fluidcomposition comprises an uncrosslinked lubricin where the uncrosslinkedlubricin represents, e.g., about 90% by weight, about 91% by weight,about 92% by weight, about 93% by weight, about 94% by weight, about 95%by weight, about 96% by weight, about 97% by weight, about 98% byweight, or about 99%, or about 100% by weight, of the total lubricinpresent in the composition. In other aspects of this embodiment, a fluidcomposition comprises an uncrosslinked lubricin where the uncrosslinkedlubricin represents, e.g., at least 90% by weight, at least 91% byweight, at least 92% by weight, at least 93% by weight, at least 94% byweight, at least 95% by weight, at least 96% by weight, at least 97% byweight, at least 98% by weight, or at least 99% by weight, of the totallubricin present in the composition. In yet other aspects of thisembodiment, a fluid composition comprises an uncrosslinked lubricinwhere the uncrosslinked lubricin represents, e.g., about 90% to about100% by weight, about 93% to about 100% by weight, about 95% to about100% by weight, or about 97% to about 100% by weight, of the totallubricin present in the composition.

In another embodiment, a fluid composition comprises an uncrosslinkedmatrix polymer that is entirely free of a crosslinked matrix polymer.

In yet another embodiment, a fluid composition comprises anuncrosslinked glycosaminoglycan that is entirely free of a crosslinkedglycosaminoglycan. In an aspect of this embodiment, a fluid compositioncomprises an uncrosslinked chondroitin sulfate polymer that is entirelyfree of a crosslinked chondroitin sulfate polymer. In another aspect ofthis embodiment, a fluid composition comprises an uncrosslinked keratansulfate polymer that is entirely free of a crosslinked keratan sulfatepolymer. In yet another aspect of this embodiment, a fluid compositioncomprises an uncrosslinked hyaluronan polymer that is entirely free of acrosslinked hyaluronan polymer.

In still another embodiment, a fluid composition comprises anuncrosslinked lubricin that is entirely free of a crosslinked lubricin.

In another embodiment, a fluid composition comprises an uncrosslinkedmatrix polymer that is essentially free of a crosslinked matrix polymer.

In yet another embodiment, a fluid composition comprises anuncrosslinked glycosaminoglycan that is essentially free of acrosslinked glycosaminoglycan. In an aspect of this embodiment, a fluidcomposition comprises an uncrosslinked chondroitin sulfate polymer thatis essentially free of a crosslinked chondroitin sulfate polymer. Inanother aspect of this embodiment, a fluid composition comprises anuncrosslinked keratan sulfate polymer that is essentially free of acrosslinked keratan sulfate polymer. In yet another aspect of thisembodiment, a fluid composition comprises an uncrosslinked hyaluronanpolymer that is essentially free of a crosslinked hyaluronan polymer.

In still another embodiment, a fluid composition comprises anuncrosslinked lubricin that is essentially free of a crosslinkedlubricin.

In another embodiment, a fluid composition comprises a crosslinkedmatrix polymer. In other aspects of this embodiment, a fluid compositioncomprises a crosslinked matrix polymer where the partially crosslinkedmatrix polymer represents, e.g., about 1% by weight, about 2% by weight,about 3% by weight, about 4% by weight, about 5% by weight, about 6% byweight, about 7% by weight, about 8% by weight, or about 9%, or about10% by weight, of the total matrix polymer present in the composition.In yet other aspects of this embodiment, a fluid composition comprises acrosslinked matrix polymer where the partially crosslinked matrixpolymer represents, e.g., at most 1% by weight, at most 2% by weight, atmost 3% by weight, at most 4% by weight, at most 5% by weight, at most6% by weight, at most 7% by weight, at most 8% by weight, at most 9% byweight, or at most 10% by weight, of the total matrix polymer present inthe composition. In still other aspects of this embodiment, a fluidcomposition comprises a crosslinked matrix polymer where the partiallycrosslinked matrix polymer represents, e.g., about 0% to about 10% byweight, about 1% to about 10% by weight, about 3% to about 10% byweight, or about 5% to about 10% by weight, of the total matrix polymerpresent in the composition.

In other aspects of this embodiment, a fluid composition comprises acrosslinked matrix polymer where the degree of crosslinking is about 1%,about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%,about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, orabout 15%. In yet other aspects of this embodiment, a fluid compositioncomprises a crosslinked matrix polymer where the degree of crosslinkingis at most 1%, at most 2%, at most 3%, at most 4%, at most 5%, at most6%, at most 7%, at most 8%, at most 9%, at most 10%, at most 11%, atmost 12%, at most 13%, at most 14%, or at most 15%. In still otheraspects of this embodiment, a fluid composition comprises a crosslinkedmatrix polymer where the degree of crosslinking is about 1% to about15%, about 2% to about 11%, about 3% to about 10%, about 1% to about 5%,about 10% to about 15%, about 11% to about 15%, about 6% to about 10%,or about 6% to about 8%.

In yet another embodiment, a fluid composition comprises a crosslinkedglycosaminoglycan. In aspect of this embodiment, a fluid compositioncomprises a crosslinked chondroitin sulfate polymer, a crosslinkedkeratan sulfate polymer, or a crosslinked hyaluronan polymer. In otheraspects of this embodiment, a fluid composition comprises a crosslinkedglycosaminoglycan where the crosslinked glycosaminoglycan represents,e.g., about 1% by weight, about 2% by weight, about 3% by weight, about4% by weight, about 5% by weight, about 6% by weight, about 7% byweight, about 8% by weight, or about 9%, or about 10% by weight, of thetotal glycosaminoglycan present in the composition. In yet other aspectsof this embodiment, a fluid composition comprises a crosslinkedglycosaminoglycan where the crosslinked glycosaminoglycan represents,e.g., at most 1% by weight, at most 2% by weight, at most 3% by weight,at most 4% by weight, at most 5% by weight, at most 6% by weight, atmost 7% by weight, at most 8% by weight, at most 9% by weight, or atmost 10% by weight, of the total glycosaminoglycan present in thecomposition. In still other aspects of this embodiment, a fluidcomposition comprises a crosslinked glycosaminoglycan where thecrosslinked glycosaminoglycan represents, e.g., about 0% to about 10% byweight, about 1% to about 10% by weight, about 3% to about 10% byweight, or about 5% to about 10% by weight, of the totalglycosaminoglycan present in the composition.

In other aspects of this embodiment, a fluid composition comprises acrosslinked glycosaminoglycan where the degree of crosslinking is about1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, orabout 15%. In yet other aspects of this embodiment, a fluid compositioncomprises a crosslinked glycosaminoglycan where the degree ofcrosslinking is at most 1%, at most 2%, at most 3%, at most 4%, at most5%, at most 6%, at most 7%, at most 8%, at most 9%, at most 10%, at most11%, at most 12%, at most 13%, at most 14%, or at most 15%. In stillother aspects of this embodiment, a fluid composition comprises acrosslinked glycosaminoglycan where the degree of crosslinking is about1% to about 15%, about 2% to about 11%, about 3% to about 10%, about 1%to about 5%, about 10% to about 15%, about 11% to about 15%, about 6% toabout 10%, or about 6% to about 8%.

In still another embodiment, a fluid composition comprises a crosslinkedlubricin. In aspects of this embodiment, a fluid composition comprises acrosslinked lubricin where the crosslinked lubricin represents, e.g.,about 1% by weight, about 2% by weight, about 3% by weight, about 4% byweight, about 5% by weight, about 6% by weight, about 7% by weight,about 8% by weight, or about 9%, or about 10% by weight, of the totallubricin present in the composition. In other aspects of thisembodiment, a fluid composition comprises a crosslinked lubricin wherethe crosslinked lubricin represents, e.g., at most 1% by weight, at most2% by weight, at most 3% by weight, at most 4% by weight, at most 5% byweight, at most 6% by weight, at most 7% by weight, at most 8% byweight, at most 9% by weight, or at most 10% by weight, of the totallubricin present in the composition. In yet other aspects of thisembodiment, a fluid composition comprises a crosslinked lubricin wherethe crosslinked lubricin represents, e.g., about 0% to about 10% byweight, about 1% to about 10% by weight, about 3% to about 10% byweight, or about 5% to about 10% by weight, of the total lubricinpresent in the composition.

In other aspects of this embodiment, a fluid composition comprises acrosslinked lubricin where the degree of crosslinking is about 1%, about2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15%.In yet other aspects of this embodiment, a fluid composition comprises acrosslinked lubricin where the degree of crosslinking is at most 1%, atmost 2%, at most 3%, at most 4%, at most 5%, at most 6%, at most 7%, atmost 8%, at most 9%, at most 10%, at most 11%, at most 12%, at most 13%,at most 14%, or at most 15%. In still other aspects of this embodiment,a fluid composition comprises a crosslinked lubricin where the degree ofcrosslinking is about 1% to about 15%, about 2% to about 11%, about 3%to about 10%, about 1% to about 5%, about 10% to about 15%, about 11% toabout 15%, about 6% to about 10%, or about 6% to about 8%.

In another embodiment, a fluid composition comprises an uncrosslinkedmatrix polymer where the uncrosslinked matrix polymer is present in anamount sufficient to improve a condition of the skin, such as, e.g.,hydration or elasticity. In aspects of this embodiment, a fluidcomposition comprises an uncrosslinked matrix polymer where theuncrosslinked matrix polymer is present at a concentration of, e.g.,about 5 mg/mL, about 6 mg/mL, about 7 mg/mL, about 8 mg/mL, about 9mg/mL, about 10 mg/mL, about 11 mg/mL, about 12 mg/mL, about 13 mg/mL,about 13.5 mg/mL, about 14 mg/mL, about 15 mg/mL, about 16 mg/mL, about17 mg/mL, about 18 mg/mL, about 19 mg/mL, or about 20 mg/mL. In otheraspects of this embodiment, a fluid composition comprises anuncrosslinked matrix polymer where the uncrosslinked matrix polymer ispresent at a concentration of, e.g., at least 1 mg/mL, at least 5 mg/mL,at least 10 mg/mL, at least 15 mg/mL, at least 20 mg/mL, or at least 25mg/mL. In yet other aspects of this embodiment, a fluid compositioncomprises an uncrosslinked matrix polymer where the uncrosslinked matrixpolymer is present at a concentration of, e.g., at most 1 mg/mL, at most5 mg/mL, at most 10 mg/mL, at most 15 mg/mL, at most 20 mg/mL, or atmost 25 mg/mL. In still other aspects of this embodiment, a fluidcomposition comprises an uncrosslinked matrix polymer where theuncrosslinked matrix polymer is present at a concentration of, e.g.,about 7.5 mg/mL to about 19.5 mg/mL, about 8.5 mg/mL to about 18.5mg/mL, about 9.5 mg/mL to about 17.5 mg/mL, about 10.5 mg/mL to about16.5 mg/mL, about 11.5 mg/mL to about 15.5 mg/mL, or about 12.5 mg/mL toabout 14.5 mg/mL.

In yet another embodiment, a fluid composition comprises anuncrosslinked glycosaminoglycan where the uncrosslinkedglycosaminoglycan is present in an amount sufficient to improve acondition of the skin, such as, e.g., hydration or elasticity. Inaspects of this embodiment, a fluid composition comprises anuncrosslinked glycosaminoglycan where the uncrosslinkedglycosaminoglycan is present at a concentration of, e.g., about 2 mg/mL,about 3 mg/mL, about 4 mg/mL, about 5 mg/mL, about 6 mg/mL, about 7mg/mL, about 8 mg/mL, about 9 mg/mL, about 10 mg/mL, about 11 mg/mL,about 12 mg/mL, about 13 mg/mL, about 13.5 mg/mL, about 14 mg/mL, about15 mg/mL, about 16 mg/mL, about 17 mg/mL, about 18 mg/mL, about 19mg/mL, or about 20 mg/mL. In other aspects of this embodiment, a fluidcomposition comprises an uncrosslinked glycosaminoglycan where theuncrosslinked glycosaminoglycan is present at a concentration of, e.g.,at least 1 mg/mL, at least 2 mg/mL, at least 3 mg/mL, at least 4 mg/mL,at least 5 mg/mL, at least 10 mg/mL, at least 15 mg/mL, at least 20mg/mL, or at least 25 mg/mL. In yet other aspects of this embodiment, afluid composition comprises an uncrosslinked glycosaminoglycan where theuncrosslinked glycosaminoglycan is present at a concentration of, e.g.,at most 1 mg/mL, at most 2 mg/mL, at most 3 mg/mL, at most 4 mg/mL, atmost 5 mg/mL, at most 10 mg/mL, at most 15 mg/mL, at most 20 mg/mL, orat most 25 mg/mL. In still other aspects of this embodiment, a fluidcomposition comprises an uncrosslinked glycosaminoglycan where theuncrosslinked glycosaminoglycan is present at a concentration of, e.g.,about 7.5 mg/mL to about 19.5 mg/mL, about 8.5 mg/mL to about 18.5mg/mL, about 9.5 mg/mL to about 17.5 mg/mL, about 10.5 mg/mL to about16.5 mg/mL, about 11.5 mg/mL to about 15.5 mg/mL, or about 12.5 mg/mL toabout 14.5 mg/mL.

In still another embodiment, a fluid composition comprises anuncrosslinked lubricin where the uncrosslinked lubricin is present in anamount sufficient to improve a condition of the skin, such as, e.g.,hydration or elasticity. In aspects of this embodiment, a fluidcomposition comprises an uncrosslinked lubricin where the uncrosslinkedlubricin is present at a concentration of, e.g., about 5 mg/mL, about 6mg/mL, about 7 mg/mL, about 8 mg/mL, about 9 mg/mL, about 10 mg/mL,about 11 mg/mL, about 12 mg/mL, about 13 mg/mL, about 13.5 mg/mL, about14 mg/mL, about 15 mg/mL, about 16 mg/mL, about 17 mg/mL, about 18mg/mL, about 19 mg/mL, or about 20 mg/mL. In other aspects of thisembodiment, a fluid composition comprises an uncrosslinked lubricinwhere the uncrosslinked lubricin is present at a concentration of, e.g.,at least 1 mg/mL, at least 5 mg/mL, at least 10 mg/mL, at least 15mg/mL, at least 20 mg/mL, or at least 25 mg/mL. In yet other aspects ofthis embodiment, a fluid composition comprises an uncrosslinked lubricinwhere the uncrosslinked lubricin is present at a concentration of, e.g.,at most 1 mg/mL, at most 5 mg/mL, at most 10 mg/mL, at most 15 mg/mL, atmost 20 mg/mL, or at most 25 mg/mL. In still other aspects of thisembodiment, a fluid composition comprises an uncrosslinked lubricinwhere the uncrosslinked lubricin is present at a concentration of, e.g.,about 7.5 mg/mL to about 19.5 mg/mL, about 8.5 mg/mL to about 18.5mg/mL, about 9.5 mg/mL to about 17.5 mg/mL, about 10.5 mg/mL to about16.5 mg/mL, about 11.5 mg/mL to about 15.5 mg/mL, or about 12.5 mg/mL toabout 14.5 mg/mL.

In a further embodiment, a fluid composition comprises a crosslinkedmatrix polymer and an uncrosslinked matrix polymer. In another aspect ofthis embodiment, a fluid composition comprises a crosslinked matrixpolymer and an uncrosslinked matrix polymer where the gel:fluid ratio issufficient to form a fluid. In other aspects of this embodiment, a fluidcomposition comprises a crosslinked matrix polymer and an uncrosslinkedmatrix polymer where the gel:fluid ratio is, e.g., about 0:100, about1:99, about 2:98, about 3:97, about 4:96, about 5:95, about 6:94, about7:93, about 8:92, about 9:91, or about 10:90. In yet other aspects ofthis embodiment, a fluid composition comprises a crosslinked matrixpolymer and an uncrosslinked matrix polymer where the gel:fluid ratiois, e.g., at most 1:99, at most 2:98, at most 3:97, at most 4:96, atmost 5:95, at most 6:94, at most 7:93, at most 8:92, at most 9:91, or atmost 10:90. In still other aspects of this embodiment, a fluidcomposition comprises a crosslinked matrix polymer and an uncrosslinkedmatrix polymer where the gel:fluid ratio is, e.g., about 0:100 to about3:97, about 0:100 to about 5:95, or about 0:100 to about 10:90.

In other aspects of this embodiment, a fluid composition comprises acrosslinked matrix polymer and an uncrosslinked matrix polymer where thegel:fluid ratio is, e.g., about 15:85, about 20:80, about 25:75, about30:70, about 35:65, about 40:60, about 45:55, about 50:50, about 55:45,about 60:40, about 65:35, about 70:30, about 75:25, about 80:20, about85:15, about 90:10, about 95:5, about 98:2, or about 100:0. In yet otheraspects of this embodiment, a fluid composition comprises a crosslinkedmatrix polymer and an uncrosslinked matrix polymer where the gel:fluidratio is, e.g., at most 15:85, at most 20:80, at most 25:75, at most30:70, at most 35:65, at most 40:60, at most 45:55, at most 50:50, atmost 55:45, at most 60:40, at most 65:35, at most 70:30, at most 75:25,at most 80:20, at most 85:15, at most 90:10, at most 95:5, at most 98:2,or at most 100:0. In still other aspects of this embodiment, a fluidcomposition comprises a crosslinked matrix polymer and an uncrosslinkedmatrix polymer where the gel:fluid ratio is, e.g., about 10:90 to about70:30, about 15:85 to about 70:30, about 10:90 to about 55:45, about80:20 to about 95:5, about 90:10 to about 100:0, about 75:25 to about100:0, or about 60:40 to about 100:0.

In another embodiment, a fluid composition comprises a substantiallyuncrosslinked hyaluronan. In aspects of this embodiment, a fluidcomposition comprises an uncrosslinked hyaluronan where theuncrosslinked hyaluronan represents, e.g., about 90% by weight, about91% by weight, about 92% by weight, about 93% by weight, about 94% byweight, about 95% by weight, about 96% by weight, about 97% by weight,about 98% by weight, or about 99%, or about 100% by weight, of the totalhyaluronan present in the composition. In other aspects of thisembodiment, a fluid composition comprises an uncrosslinked hyaluronanwhere the uncrosslinked hyaluronan represents, e.g., at least 90% byweight, at least 91% by weight, at least 92% by weight, at least 93% byweight, at least 94% by weight, at least 95% by weight, at least 96% byweight, at least 97% by weight, at least 98% by weight, or at least 99%by weight, of the total hyaluronan present in the composition. In yetother aspects of this embodiment, a fluid composition comprises anuncrosslinked hyaluronan where the uncrosslinked hyaluronan represents,e.g., about 90% to about 100% by weight, about 93% to about 100% byweight, about 95% to about 100% by weight, or about 97% to about 100% byweight, of the total hyaluronan present in the composition.

In yet another embodiment, a fluid composition comprises anuncrosslinked hyaluronan where the uncrosslinked hyaluronan is presentin an amount sufficient to improve a condition of the skin, such as,e.g., hydration or elasticity. In aspects of this embodiment, a fluidcomposition comprises an uncrosslinked hyaluronan where theuncrosslinked hyaluronan is present at a concentration of, e.g., about 5mg/mL, about 6 mg/mL, about 7 mg/mL, about 8 mg/mL, about 9 mg/mL, about10 mg/mL, about 11 mg/mL, about 12 mg/mL, about 13 mg/mL, about 13.5mg/mL, about 14 mg/mL, about 15 mg/mL, about 16 mg/mL, about 17 mg/mL,about 18 mg/mL, about 19 mg/mL, or about 20 mg/mL. In other aspects ofthis embodiment, a fluid composition comprises an uncrosslinkedhyaluronan where the uncrosslinked hyaluronan is present at aconcentration of, e.g., at least 1 mg/mL, at least 5 mg/mL, at least 10mg/mL, at least 15 mg/mL, at least 20 mg/mL, or at least 25 mg/mL. Inyet other aspects of this embodiment, a fluid composition comprises anuncrosslinked hyaluronan where the uncrosslinked hyaluronan is presentat a concentration of, e.g., at most 1 mg/mL, at most 5 mg/mL, at most10 mg/mL, at most 15 mg/mL, at most 20 mg/mL, or at most 25 mg/mL. Instill other aspects of this embodiment, a fluid composition comprises anuncrosslinked hyaluronan where the uncrosslinked hyaluronan is presentat a concentration of, e.g., about 7.5 mg/mL to about 19.5 mg/mL, about8.5 mg/mL to about 18.5 mg/mL, about 9.5 mg/mL to about 17.5 mg/mL,about 10.5 mg/mL to about 16.5 mg/mL, about 11.5 mg/mL to about 15.5mg/mL, or about 12.5 mg/mL to about 14.5 mg/mL.

In other aspects of this embodiment, a fluid composition comprises anuncrosslinked hyaluronan where the uncrosslinked hyaluronan has a meanmolecular weight of, e.g., about 1,000,000 Da, about 1,500,000 Da, about2,000,000 Da, about 2,500,000 Da, about 3,000,000 Da, about 3,500,000Da, about 4,000,000 Da, about 4,500,000 Da, or about 5,000,000 Da. Inyet other aspects of this embodiment, a fluid composition comprises anuncrosslinked hyaluronan where the uncrosslinked hyaluronan has a meanmolecular weight of, e.g., at least 1,000,000 Da, at least 1,500,000 Da,at least 2,000,000 Da, at least 2,500,000 Da, at least 3,000,000 Da, atleast 3,500,000 Da, at least 4,000,000 Da, at least 4,500,000 Da, or atleast 5,000,000 Da. In still other aspects of this embodiment, a fluidcomposition comprises an uncrosslinked hyaluronan where theuncrosslinked hyaluronan has a mean molecular weight of, e.g., about1,000,000 Da to about 5,000,000 Da, about 1,500,000 Da to about5,000,000 Da, about 2,000,000 Da to about 5,000,000 Da, about 2,500,000Da to about 5,000,000 Da, about 2,000,000 Da to about 3,000,000 Da,about 2,500,000 Da to about 3,500,000 Da, or about 2,000,000 Da to about4,000,000 Da. In further aspects, a fluid composition comprises anuncrosslinked hyaluronan where the uncrosslinked hyaluronan has a meanmolecular weight of, e.g., greater than 2,000,000 Da and less than about3,000,000 Da, greater than 2,000,000 Da and less than about 3,500,000Da, greater than 2,000,000 Da and less than about 4,000,000 Da, greaterthan 2,000,000 Da and less than about 4,500,000 Da, greater than2,000,000 Da and less than about 5,000,000 Da.

In another embodiment, a fluid composition comprises an uncrosslinkedhyaluronan where the uncrosslinked hyaluronan comprises a combination ofboth high molecular weight hyaluronan and low molecular weighthyaluronan, in various ratios. As used herein, the term “high molecularweight hyaluronan” refers to a hyaluronan polymer that has a molecularweight of 1,000,000 Da or greater. Non-limiting examples of a highmolecular weight hyaluronan include a hyaluronan of about 1,500,000 Da,a hyaluronan of about 2,000,000 Da, a hyaluronan of about 2,500,000 Da,a hyaluronan of about 3,000,000 Da, a hyaluronan of about 3,500,000 Da,a hyaluronan of about 4,000,000 Da, a hyaluronan of about 4,500,000 Da,and a hyaluronan of about 5,000,000 Da. As used herein, the term “lowmolecular weight hyaluronan” refers to a hyaluronan polymer that has amolecular weight of less than 1,000,000 Da. Non-limiting examples of alow molecular weight hyaluronan include a hyaluronan of about 200,000Da, a hyaluronan of about 300,000 Da, a hyaluronan of about 400,000 Da,a hyaluronan of about 500,000 Da, a hyaluronan of about 600,000 Da, ahyaluronan of about 700,000 Da, a hyaluronan of about 800,000 Da, and ahyaluronan of about 900,000 Da.

Thus, in an embodiment, a fluid composition comprises an uncrosslinkedhyaluronan where the uncrosslinked hyaluronan comprises a combination ofboth high molecular weight hyaluronan and low molecular weighthyaluronan in a ratio of about 20:1, about 15:1, about 10:1, about 5:1,about 1:1, about 1:5 about 1:10, about 1:15, or about 1:20.

In still another embodiment, a fluid composition comprises a crosslinkedhyaluronan. In aspects of this embodiment, a fluid composition comprisesa crosslinked hyaluronan where the crosslinked hyaluronan represents,e.g., about 1% by weight, about 2% by weight, about 3% by weight, about4% by weight, about 5% by weight, about 6% by weight, about 7% byweight, about 8% by weight, or about 9%, or about 10% by weight, of thetotal hyaluronan present in the composition. In other aspects of thisembodiment, a fluid composition comprises a crosslinked hyaluronan wherethe crosslinked hyaluronan represents, e.g., at most 1% by weight, atmost 2% by weight, at most 3% by weight, at most 4% by weight, at most5% by weight, at most 6% by weight, at most 7% by weight, at most 8% byweight, at most 9% by weight, or at most 10% by weight, of the totalhyaluronan present in the composition. In yet other aspects of thisembodiment, a fluid composition comprises a crosslinked hyaluronan wherethe crosslinked hyaluronan represents, e.g., about 0% to about 10% byweight, about 1% to about 10% by weight, about 3% to about 10% byweight, or about 5% to about 10% by weight, of the total hyaluronanpresent in the composition.

In other aspects of this embodiment, a fluid composition comprises acrosslinked hyaluronan where the degree of crosslinking is about 1%,about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%,about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, orabout 15%. In yet other aspects of this embodiment, a fluid compositioncomprises a crosslinked hyaluronan where the degree of crosslinking isat most 1%, at most 2%, at most 3%, at most 4%, at most 5%, at most 6%,at most 7%, at most 8%, at most 9%, at most 10%, at most 11%, at most12%, at most 13%, at most 14%, or at most 15%. In still other aspects ofthis embodiment, a fluid composition comprises a crosslinked hyaluronanwhere the degree of crosslinking is about 1% to about 15%, about 2% toabout 11%, about 3% to about 10%, about 1% to about 5%, about 10% toabout 15%, about 11% to about 15%, about 6% to about 10%, or about 6% toabout 8%.

In other aspects of this embodiment, a fluid composition comprises acrosslinked hyaluronan where the crosslinked hyaluronan has a meanmolecular weight of, e.g., about 1,000,000 Da, about 1,500,000 Da, about2,000,000 Da, about 2,500,000 Da, about 3,000,000 Da, about 3,500,000Da, about 4,000,000 Da, about 4,500,000 Da, or about 5,000,000 Da. Inyet other aspects of this embodiment, a fluid composition comprises acrosslinked hyaluronan where the crosslinked hyaluronan has a meanmolecular weight of, e.g., at least 1,000,000 Da, at least 1,500,000 Da,at least 2,000,000 Da, at least 2,500,000 Da, at least 3,000,000 Da, atleast 3,500,000 Da, at least 4,000,000 Da, at least 4,500,000 Da, or atleast 5,000,000 Da. In still other aspects of this embodiment, a fluidcomposition comprises a crosslinked hyaluronan where the crosslinkedhyaluronan has a mean molecular weight of, e.g., about 1,000,000 Da toabout 5,000,000 Da, about 1,500,000 Da to about 5,000,000 Da, about2,000,000 Da to about 5,000,000 Da, about 2,500,000 Da to about5,000,000 Da, about 2,000,000 Da to about 3,000,000 Da, about 2,500,000Da to about 3,500,000 Da, or about 2,000,000 Da to about 4,000,000 Da.

In a further embodiment, a fluid composition comprises a crosslinkedhyaluronan and an uncrosslinked hyaluronan. In an aspect of thisembodiment, a fluid composition comprises a crosslinked hyaluronan andan uncrosslinked hyaluronan where the gel:fluid ratio is sufficient toform a fluid. In other aspects of this embodiment, a fluid compositioncomprises a crosslinked hyaluronan and an uncrosslinked hyaluronan wherethe gel:fluid ratio is, e.g., about 0:100, about 1:99, about 2:98, about3:97, about 4:96, about 5:95, about 6:94, about 7:93, about 8:92, about9:91, or about 10:90. In yet other aspects of this embodiment, a fluidcomposition comprises a crosslinked hyaluronan and an uncrosslinkedhyaluronan where the gel:fluid ratio is, e.g., at most 1:99, at most2:98, at most 3:97, at most 4:96, at most 5:95, at most 6:94, at most7:93, at most 8:92, at most 9:91, or at most 10:90. In still otheraspects of this embodiment, a fluid composition comprises a crosslinkedhyaluronan and an uncrosslinked hyaluronan where the gel:fluid ratio is,e.g., about 0:100 to about 3:97, about 0:100 to about 5:95, or about0:100 to about 10:90.

In other aspects of this embodiment, a fluid composition comprises acrosslinked hyaluronan and an uncrosslinked hyaluronan where thegel:fluid ratio is, e.g., about 15:85, about 20:80, about 25:75, about30:70, about 35:65, about 40:60, about 45:55, about 50:50, about 55:45,about 60:40, about 65:35, about 70:30, about 75:25, about 80:20, about85:15, about 90:10, about 95:5, about 98:2, or about 100:0. In yet otheraspects of this embodiment, a fluid composition comprises a crosslinkedhyaluronan and an uncrosslinked hyaluronan where the gel:fluid ratio is,e.g., at most 15:85, at most 20:80, at most 25:75, at most 30:70, atmost 35:65, at most 40:60, at most 45:55, at most 50:50, at most 55:45,at most 60:40, at most 65:35, at most 70:30, at most 75:25, at most80:20, at most 85:15, at most 90:10, at most 95:5, at most 98:2, or atmost 100:0. In still other aspects of this embodiment, a fluidcomposition comprises a crosslinked hyaluronan and an uncrosslinkedhyaluronan where the gel:fluid ratio is, e.g., about 10:90 to about70:30, about 15:85 to about 70:30, about 10:90 to about 55:45, about80:20 to about 95:5, about 90:10 to about 100:0, about 75:25 to about100:0, or about 60:40 to about 100:0.

Aspects of the present specification provide, in part, a fluidcomposition comprising a stabilizing component. As used herein, the term“stabilizing component” refers to a molecule that reduces or preventsthe degradation of a matrix polymer disclosed in the presentspecification. The stabilizing component can reduce or prevent enzymaticdegradation and/or reduce or prevent chemical degradation. Non-limitingexamples of stabilizing components include a polyol and a flavonoid.

Aspects of the present specification provide, in part, a fluidcomposition comprising a polyol. As used herein, the term “polyol” issynonymous with “sugar alcohol,” “polyhydric alcohol,” and “polyalcohol”and refers to a hydrogenated form of carbohydrate, whose carbonyl group(aldehyde or ketone, reducing sugar) has been reduced to a primary orsecondary hydroxyl group (hence the alcohol), such as, e.g., mannitolfrom mannose, xylitol from xylose, and lactitol from lactulose. Polyolshave the general formula H(HCHO)_(n)+1 H. Both monosaccharides anddisaccharides can form polyols; however, polyols derived fromdisaccharides are not entirely hydrogenated because only one aldehydegroup is available for reduction. A fluid composition disclosed in thepresent specification may comprise a single polyol, or a plurality ofpolyols.

A fluid composition disclosed in the present specification comprises apolyol that is pharmaceutically acceptable. As used herein, the term“pharmaceutically acceptable” means any molecular entity or compositionthat does not produce an adverse, allergic or other untoward or unwantedreaction when administered to a mammal. It is known in the art that thetwo-carbon polyol, glycol, is not pharmaceutically acceptable becausethis polyol is toxic to mammals. It is also known in the art thatpolyols comprising three or more carbon atoms are typicallypharmaceutically acceptable. As such, polyols comprising three or morecarbon atoms are generally useful in making the compositions disclosedin the present specification.

A fluid composition disclosed in the present specification comprises apolyol in an amount sufficient to protect uncrosslinkedglycosaminoglycans, like hyaluronan, from degradation, such as enzymaticdegradation and chemical degradation. One primary means of suchglycosaminoglycan degradation is chemical breakdown from exposure tofree radicals, such as, e.g., OH. radicals. Free radicals are atoms,molecules, or ions with unpaired electrons on an open shellconfiguration. The unpaired electrons cause them to be highly chemicallyreactive.

Free radicals play an important role in a number of biologicalprocesses, some of which are necessary for life, such as theintracellular killing of bacteria by phagocytic cells such asgranulocytes and macrophages. Free radicals have also been implicated incertain cell signaling processes, called redox signaling. The two mostimportant oxygen-centered free radicals are superoxide and hydroxylradical. They are derived from molecular oxygen under reducingconditions. For example, the superoxide anion (O₂ ⁻) can capture anotherelectron to form the peroxide ion (O₂ ²⁻), which in turn may react withtwo H⁺ protons to form hydrogen peroxide (H₂O₂). The degradation ofhydrogen peroxide in the presence of transition metals in their reducedform (Fenton's reaction) leads to the formation of the hydroxyl radical(OH.). A free radical formed in this way can give rise to a series ofreactions leading to the formation of different species of activeoxygen. Table 2 describes the principal active species of oxygen.

TABLE 2 Principal species of active oxygen Chemical Name Symbol CommentsSuperoxide O₂ ⁻ Superoxide anions are formed by reduction of molecularanion oxygen. They are minimally reactive in aqueous media, whichenables them to migrate quite a long way from their site of production.Superoxide anions have a weak oxidative action but are capable ofgenerating more reactive radicals. Hydrogen H₂O₂ Hydrogen peroxides areformed either by bivalent reduction peroxide of molecular oxygen, or bydismutation of the superoxide anion. The absence of electric charges ontheir surface makes them very lipophilic and minimally reactive inaqueous media. The degradation of hydrogen peroxides (Fenton's reaction)produces very reactive radicals called hydroxyl radicals. Hydroxylradical OH^(•) Hydroxyl radicals are formed by degradation of hydrogenperoxide in the presence of transition metals in their reduced form(Fenton's reaction). Hydroxyl radicals are very reactive. Theirhalf-life is on the order of 10⁻⁹ s. Peroxyl radical ROO^(•) Peroxylradicals are formed by addition of molecular oxygen to free carbonradicals. They are minimally reactive. Organic ROOH Organichydroperoxides are the protonated form of peroxyl hydroperoxideradicals. They are very reactive and decompose anew into peroxylradicals and alcoxyl radicals. Alcoxyl radical RO^(•) Alcoxyl radicalsare formed during the degradation of organic peroxides. They are veryreactive. Their half-life is on the order of 10⁻⁶ s. Nitric oxide NO^(•)Nitric oxides are synthesized from arginine (an amino acid) via theaction of nitric oxide synthetase. They interact with hydroxyl radicalsto form peroxynitrites. Nitric oxide is a neuromediator and can also beused by immune cells to destroy microbes or dangerous cells.Peroxynitrite ONOO⁻ Peroxynitrites are capable of oxidizing certainsubstances such as methionine (an amino acid serving as a constituent ofproteins and enzymes) or of reacting with SOD (Cf. § 1.3.2.a) thereby“nitrating” tyrosine (another very important amino acid) Nitrosylradical ONOOH Degradation of nitrosyl radicals leads to the formation ofhydroxyl radicals. Oxidizing power : OH^(•) > RO^(•) > ROO^(•) > NO^(•)

However, because of their reactivity, these same free radicals canparticipate in unwanted side reactions resulting in cell damage. Forpurposes of the present specification, OH. radicals interact with thehydrogen located on the carbon adjacent to the carboxyl group in theglucuronic ring of hyaluronan and other glycosaminoglycans, therebyremoving it. This removal causes splitting of the glycoside bond andhence depolymerization of the polymer.

In general, the crosslinking observed in the hyaluronan and other matrixpolymers used in dermal fillers protect these polymers against thechemical breakdown of free radical species of oxygen. This is becausethe bonds formed during crosslinking mask the hydrogen attached by thefree radicals. However, the uncrosslinked glycosaminoglycans disclosedin the present specification is afforded no such protection. The presentspecification discloses an alternative means of protecting matrixpolymers from oxidative degradation of free radicals. It has beendetermined that polyols disclosed in the present specification act asstabilizing agents that can neutralize free radicals of active oxygen.As a stabilizing, a polyol is stabilizing component that protects matrixpolymers like hyaluronan against the effects of oxidative stress andlimiting the degradation of the fluid compositions disclosed in thepresent specification.

Thus, any polyol is useful in making the compositions disclosed in thepresent specification, with the proviso that the polyol is non-toxic toa mammal and the polyol protects uncrosslinked matrix polymers likehyaluronan from degradation. Non-limiting examples of polyols include,glycerol, erythritol, threitol, arabitol, erythritol, ribitol, xylitol,galactitol (or dulcitol), glucitol (or sorbitol), iditol, inositol,mannitol, isomalt, lactitol, maltitol, and polyglycitol. Othernon-limiting examples of polyols can be found in, e.g., PharmaceuticalDosage Forms and Drug Delivery Systems (Howard C. Ansel et al., eds.,Lippincott Williams & Wilkins Publishers, 7^(th) ed. 1999); Remington:The Science and Practice of Pharmacy (Alfonso R. Gennaro ed.,Lippincott, Williams & Wilkins, 20^(th) ed. 2000); Goodman & Gilman'sThe Pharmacological Basis of Therapeutics (Joel G. Hardman et al., eds.,McGraw-Hill Professional, 10^(th) ed. 2001); and Handbook ofPharmaceutical Excipients (Raymond C. Rowe et al., APhA Publications,4^(th) edition 2003), each of which is hereby incorporated by referencein its entirety.

Thus in an embodiment, a fluid composition comprises a pharmaceuticallyacceptable polyol that can reduce or prevent degradation of a matrixpolymer. In aspects of this embodiment, a fluid composition comprises apharmaceutically acceptable three-carbon polyol, a pharmaceuticallyacceptable four-carbon polyol, a pharmaceutically acceptable five-carbonpolyol, a pharmaceutically acceptable six-carbon polyol, apharmaceutically acceptable seven-carbon polyol, a pharmaceuticallyacceptable eight-carbon polyol, a pharmaceutically acceptablenine-carbon polyol, a pharmaceutically acceptable ten-carbon polyol, apharmaceutically acceptable eleven-carbon polyol, or a pharmaceuticallyacceptable twelve-carbon polyol. In other aspects of this embodiment, afluid composition comprises glycerol, erythritol, threitol, arabitol,erythritol, ribitol, xylitol, galactitol (or dulcitol), glucitol (orsorbitol), iditol, inositol, mannitol, isomalt, lactitol, maltitol, orpolyglycitol.

In another embodiment, a fluid composition comprises a single polyolthat can reduce or prevent degradation of a matrix polymer. In yetanother embodiment, a fluid composition comprises a plurality ofpolyols, each of which can reduce or prevent the degradation of a matrixpolymer. In aspects of this embodiment, a fluid composition comprisesone or more polyols, two or more polyols, three or more polyols, four ormore polyols, or five or more polyols. In other aspects of thisembodiment, a fluid composition comprises one to five polyols, two tofive polyols, three to five polyols, two to four polyols, two to fivepolyols, or three to five polyols.

In yet another embodiment, a fluid composition comprises a polyol in anamount sufficient to reduce or prevent degradation of a matrix polymer.In aspects of this embodiment, a fluid composition comprises a polyol inan amount of, e.g., about 0.1% (w/v) of the composition, about 0.2%(w/v) of the composition, about 0.3% (w/v) of the composition, about0.4% (w/v) of the composition, about 0.5% (w/v) of the composition,about 0.6% (w/v) of the composition, about 0.7% (w/v) of thecomposition, about 0.8% (w/v) of the composition, about 0.9% (w/v) ofthe composition, about 1.0% (w/v) of the composition, about 2.0% (w/v)of the composition, about 3.0% (w/v) of the composition, about 4.0%(w/v) of the composition, about 5.0% (w/v) of the composition, about6.0% (w/v) of the composition, about 7.0% (w/v) of the composition,about 8.0% (w/v) of the composition, about 9.0% (w/v) of thecomposition, or about 10% (w/v) of the composition. In other aspects, afluid composition comprises a polyol in an amount of, e.g., at least0.1% (w/v) of the composition, at least 0.2% (w/v) of the composition,at least 0.3% (w/v) of the composition, at least 0.4% (w/v) of thecomposition, at least 0.5% (w/v) of the composition, at least 0.6% (w/v)of the composition, at least 0.7% (w/v) of the composition, at least0.8% (w/v) of the composition, at least 0.9% (w/v) of the composition,at least 1.0% (w/v) of the composition, at least 2.0% (w/v) of thecomposition, at least 3.0% (w/v) of the composition, at least 4.0% (w/v)of the composition, at least 5.0% (w/v) of the composition, at least6.0% (w/v) of the composition, at least 7.0% (w/v) of the composition,at least 8.0% (w/v) of the composition, at least 9.0% (w/v) of thecomposition, or at least 10% (w/v) of the composition. In yet otheraspects, a fluid composition comprises a polyol in an amount of, e.g.,at most 0.1% (w/v) of the composition, at most 0.2% (w/v) of thecomposition, at most 0.3% (w/v) of the composition, at most 0.4% (w/v)of the composition, at most 0.5% (w/v) of the composition, at most 0.6%(w/v) of the composition, at most 0.7% (w/v) of the composition, at most0.8% (w/v) of the composition, at most 0.9% (w/v) of the composition, atmost 1.0% (w/v) of the composition, at most 2.0% (w/v) of thecomposition, at most 3.0% (w/v) of the composition, at most 4.0% (w/v)of the composition, at most 5.0% (w/v) of the composition, at most 6.0%(w/v) of the composition, at most 7.0% (w/v) of the composition, at most8.0% (w/v) of the composition, at most 9.0% (w/v) of the composition, orat most 10% (w/v) of the composition. In still other aspects, a fluidcomposition comprises a polyol in an amount of, e.g., about 0.1% (w/v)to about 1.0% (w/v) of the composition, about 0.1% (w/v) to about 2.0%(w/v) of the composition, about 0.1% (w/v) to about 3.0% (w/v) of thecomposition, about 0.1% (w/v) to about 4.0% (w/v) of the composition,about 0.1% (w/v) to about 5.0% (w/v) of the composition, about 0.2%(w/v) to about 0.9% (w/v) of the composition, about 0.2% (w/v) to about1.0% (w/v) of the composition, about 0.2% (w/v) to about 2.0% (w/v) ofthe composition, about 0.5% (w/v) to about 1.0% (w/v) of thecomposition, or about 0.5% (w/v) to about 2.0% (w/v) of the composition.

In other aspects of this embodiment, a fluid composition comprises apolyol is present at a concentration of, e.g., about 0.01 mg/mL, about0.02 mg/mL, about 0.03 mg/mL, about 0.04 mg/mL, about 0.05 mg/mL, about0.06 mg/mL, about 0.07 mg/mL, about 0.08 mg/mL, about 0.09 mg/mL, about0.1 mg/mL, about 0.2 mg/mL, about 0.3 mg/mL, about 0.4 mg/mL, about 0.5mg/mL, about 0.6 mg/mL, about 0.7 mg/mL, about 0.8 mg/mL, about 0.9mg/mL, about 1.0 mg/mL, about 2.0 mg/mL, about 3.0 mg/mL, about 4.0mg/mL, about 5.0 mg/mL, about 6.0 mg/mL, about 7.0 mg/mL, about 8.0mg/mL, about 9.0 mg/mL, or about 10 mg/mL. In yet other aspects of thisembodiment, a fluid composition comprises a polyol present at aconcentration of, e.g., at least 0.01 mg/mL, at least 0.02 mg/mL, atleast 0.03 mg/mL, at least 0.04 mg/mL, at least 0.05 mg/mL, at least0.06 mg/mL, at least 0.07 mg/mL, at least 0.08 mg/mL, at least 0.09mg/mL, at least 0.1 mg/mL, at least 0.2 mg/mL, at least 0.3 mg/mL, atleast 0.4 mg/mL, at least 0.5 mg/mL, at least 0.6 mg/mL, at least 0.7mg/mL, at least 0.8 mg/mL, at least 0.9 mg/mL, at least 1.0 mg/mL, atleast 2.0 mg/mL, at least 3.0 mg/mL, at least 4.0 mg/mL, at least 5.0mg/mL, at least 6.0 mg/mL, at least 7.0 mg/mL, at least 8.0 mg/mL, atleast 9.0 mg/mL, or at least 10 mg/mL. In still other aspects of thisembodiment, a fluid composition comprises a polyol present at aconcentration of, e.g., at most 0.01 mg/mL, at most 0.02 mg/mL, at most0.03 mg/mL, at most 0.04 mg/mL, at most 0.05 mg/mL, at most 0.06 mg/mL,at most 0.07 mg/mL, at most 0.08 mg/mL, at most 0.09 mg/mL, at most 0.1mg/mL, at most 0.2 mg/mL, at most 0.3 mg/mL, at most 0.4 mg/mL, at most0.5 mg/mL, at most 0.6 mg/mL, at most 0.7 mg/mL, at most 0.8 mg/mL, atmost 0.9 mg/mL, at most 1.0 mg/mL, at most 2.0 mg/mL, at most 3.0 mg/mL,at most 4.0 mg/mL, at most 5.0 mg/mL, at most 6.0 mg/mL, at most 7.0mg/mL, at most 8.0 mg/mL, at most 9.0 mg/mL, or at most 10 mg/mL. Infurther aspects, a fluid composition comprises a polyol present at aconcentration of, e.g., about 0.01 mg/mL to about 0.7 mg/mL, about 0.06mg/mL to about 0.7 mg/mL, about 0.01 mg/mL to about 1.0 mg/mL, about0.05 mg/mL to about 1.0 mg/mL, about 0.06 mg/mL to about 1.0 mg/mL,about 0.1 mg/mL to about 1.0 mg/mL, about 0.1 mg/mL to about 2.0 mg/mL,about 0.1 mg/mL to about 3.0 mg/mL, about 0.1 mg/mL to about 4.0 mg/mL,about 0.1 mg/mL to about 5.0 mg/mL, about 0.2 mg/mL to about 0.9 mg/mL,about 0.2 mg/mL to about 1.0 mg/mL, about 0.2 mg/mL to about 2.0 mg/mL,about 0.5 mg/mL to about 1.0 mg/mL, or about 0.5 mg/mL to about 2.0mg/mL.

Aspects of the present specification provide, in part, a fluidcomposition that can optionally comprise or not comprise a flavonoid(Table 3). A flavonoid (or bioflavonoid) refers to the class ofpolyphenolic ketone-containing and non-ketone-containing secondarymetabolites found in plants that are well known to have diversebeneficial biochemical and antioxidant effects. Non-limiting examples offlavonoids include C-methylated flavonoids, O-methylated flavonoids,isoflavonoids, neoflavonoids, flavonolignans, furanoflavonoids,pyranoflavonoids, methylenedioxyflavonoids, prenylated flavonoids,aurones, flavones, flavonols, flavanones, flavanonols, flavan-3-ols,flavan-4-ols, leucoanthocyanidin (flavan-3,4-diols), anthocyanidins, andtannins. It is understood that these and other substances known in theart of pharmacology can be included in a fluid composition disclosed inthe present specification. See for example, Remington's PharmaceuticalSciences Mac Publishing Company, Easton, Pa. 16^(th) Edition 1980.

Aurones are compounds derived from 2-benzylidene-1-benzofuran-3-one.Non-limiting examples of aurones include 4,5,6-trihydroxy-aurone,aureusidin, hispidol, leptosidin, maritimetin, and sulfuretin.

Three major classes of ketone-containing flavonoids are flavones,compounds derived from 2-phenylchromen-4-one (2-phenyl-1,4-benzopyrone);isoflavones, compounds derived from 3-phenylchromen-4-one(3-phenyl-1,4-benzopyrone); and neoflavones, compounds derived from4-phenylcoumarin (4-phenyl-1,2-benzopyrone)(Table 3). Flavones arethemselves divided into four groups based on the presence or absence of3-hydroxyl 2,3-dihydro functional groups: flavones, compounds derivedfrom 2-phenylchromen-4-one lack both functional groups; flavonols(3-hydroxyflavone), compounds derived from3-hydroxy-2-phenylchromen-4-one have the 3-hydroxyl group, but lack the2,3-dihydro group; flavanones, compounds derived from2,3-dihydro-2-phenylchromen-4-one have the 2,3-dihydro group, but lackthe 3-hydroxyl group; and flavanonols (3-hydroxyflavanone or2,3-dihydroflavonol), compounds derived from3-hydroxy-2,3-dihydro-2-phenylchromen-4-one have both functional groups.

Non-limiting examples of flavones include acacetin, apiin, apigenin,apigetrin, artoindonesianin P, baicalein, baicalin, chrysin, cynaroside,diosmetin, diosmin, eupatilin, flavoxate, 6-hydroxyflavone, genkwanin,hidrosmin, luteolin, nepetin, nepitrin (nepetin 7-glucoside), nobiletin,orientin (isoorientin), oroxindin, oroxylin A, rhoifolin, scutellarein,scutellarin, tangeritin, techtochrysin, tetuin, tricin,veronicastroside, vitexin (isovitexin), and wogonin. Non-limitingexamples of flavonols include 3-hydroxyflavone, azaleatin, fisetin,galangin, gossypetin, kaempferide, kaempferol, isorhamnetin, morin,myricetin, natsudaidain, pachypodol, quercetin, rhamnazin, rhamnetin,and sophorin. Non-limiting examples of flavanones include butin,eriodictyol, hesperetin, hesperidin, homoeriodictyol, isosakuranetin,naringenin, naringin, pinocembrin, poncirin, sakuranetin, sakuranin, andsterubin. Non-limiting examples of flavanonols include taxifolin(dihydroquercetin), and aromadedrin (dihydrokaempferol).

Isoflavonoids include isoflavones and isoflavanes (Table 3).Non-limiting examples of isoflavonoids include alpinumisoflavone,anagyroidisoflavone A and B, calycosin, daidzein, daidzin, derrubone,di-O-methylalpinumisoflavone, formononetin, genistein, genistin,glycitein, ipriflavone, irigenin, iridin, irilone,4′-methylalpinumisoflavone, 5-O-methylgenistein, luteone, ononin,orobol, pratensein, prunetin, pseudobaptigenin, psi-tectorigenin,puerarin, retusin, tectoridin, tectorigenin, and wighteone.

Neoflavonoids include 4-arylcoumarins (neoflavones), 4-arylchromanes,dalbergiones and dalbergiquinols (Table 3). Neoflavones are compoundsderived from 4-phenylcoumarin (or 4-Aryl-coumarin); neoflavenescompounds derived from 4-phenylchromen. Non-limiting examples ofneoflavonoids include calophyllolide, coutareagenin, dalbergichromene,dalbergin, and nivetin.

Non-ketone-containing flavonoids, include flavan-3-ols and catechins.Flavan-3-ols (flavanols) are a class of flavonoids derived from2-phenyl-3,4-dihydro-2H-chromen-3-ol skeleton. Catechin possesses twobenzene rings (called the A- and B-rings) and a dihydropyran heterocycle(the C-ring) with an hydroxyl group on carbon 3. The A ring is similarto a resorcinol moiety while the B ring is similar to a catechol moiety.There are two chiral centers on the molecule on carbons 2 and 3. It hastherefore four diastereoisomers. Two of the isomers are in transconfiguration and are called catechin and the other two are in cisconfiguration and are called epicatechin. Non-limiting examples ofnon-ketone-containing flavonoids include afzelechin, arthromerin A,arthromerin B, catechin, epicatechin, epigallocatechin, epicatechingallate, epigallocatechin gallate, epigallocatechin gallate,epiafzelechin, fisetinidol, gallocatechin, gallocatechin gallate,guibourtinidol, meciadanol (3-O-methylcatechin), mesquitol,robinetinidol, and thearubigin.

Flavan-4-ols (3-deoxyflavonoids) are flavone-derived alcohols derivedfrom 2-phenylchroman-4-ol. Non-limiting examples of flavan-4-ols includeapiforol and luteoforol.

Leucoanthocyanidin (flavan-3,4-diols) are compounds derived from2-phenyl-3,4-dihydro-2H-chromene-3,4-diol. Non-limiting examples offlavan-3,4-diols include leucocyanidin, leucodelphinidin, leucomalvidin,leucopelargonidin, leucopeonidin, leucorobinetinidin, and melacacidin.

Anthocyanidins are compounds derived from 2-phenylchromenylium.Non-limiting examples of anthocyanidins include antirrhinin,apigeninidin, aurantinidin, capensinidin, chrysanthenin, columnidin,commelinin, cyanidin, 6-hydroxycyanidin, cyanidin-3-(di-p-coumarylglucoside)-5-glucoside, cyanosalvian in, delphinidin, diosmetinidin, europinidin, fisetinidin, gesneridin, guibourtinidin,hirsutidin, luteolinidin, malvidin, 5-desoxy-malvidin, malvin,myrtillin, oenin, peonidin, 5-desoxy-peonidin, pelargonidin, petunidin,primulin, protocyanin, protodelphin, pulchellidin, pulchellidin3-glucoside, pulchellidin 3-rhamnoside, robinetinidin, rosinidin,tricetinidin, tulipanin, and violdelphin.

Tannins are compounds derived from 2-phenylchromenylium. There are threemajor classes of tannins: hydrolyzable tannins; non-hydrolyzable tannins(condensed tannins; proanthocyanidins); and pseudotannins.

Hydrolyzable tannins are themselves divided into four groups: oligomertannins including aglycone tannins and glycoside tannins; ellagitannins;gallotannins, and unclassified tannins. Non-limiting examples ofaglycone tannins include ellagic acid, gallagic acid, and gallic acid.Non-limiting examples of glycoside tannins include glucose, quinic acid,and shikimic acid. Non-limiting examples of ellagitannins includecastalagin (vescalagin), castalin, casuarictin, casuariin, casuarinin,cornusiin E, grandinin, pedunculagin, punicacortein C, punigluconin,punicalagin, punicalagin alpha, punicalin, 2-O-galloyl-punicalin,stachyurin, strictinin, and tellimagrandin II. Non-limiting examples ofgallotannins include corilagin, galloyl glucose, digalloyl glucose,trigalloyl glucose, tetragalloyl glucose, pentagalloyl glucose,hexagalloyl glucose, heptagalloyl glucose, octagalloyl glucose, andtannic acid. Non-limiting examples of unclassified tannins includeacutissimin A, acutissimin B, chebulagic acid, chebulinic acid,cinnamtannin B1, combreglutinin, geraniin, granatin B, roburin A,roburin B, roburin C, roburin D, roburin E, stachyurin, tercatin,terflavins A, terflavins B, tergallagin, vescalin,1,3,4-tri-O-galloylquinic acid, 3,5-di-O-galloyl-shikimic acid, and3,4,5-tri-O-galloylshikimic acid.

Condensed tannins (proanthocyanidins) are essentially polymer chains offlavonoids such as catechins. Non-limiting examples of condensed tanninsinclude proanthocyanidin, prodelphinidin, profisetinidin,proguibourtinidin, and prorobinetidin.

TABLE 3 Flavonoids Flavonoids Base compound Examples Aurones2-benzylidene-1-benzofuran-3-one 4,5,6-trihydroxy-aurone, aureusidin,hispidol, leptosidin, maritimetin, and sulfuretin Flavones2-phenylchromen-4-one acacetin, apiin, apigenin, apigetrin,artoindonesianin P, baicalein, baicalin, chrysin, cynaroside, diosmetin,diosmin, eupatilin, flavoxate, 6-hydroxyflavone, genkwanin, hidrosmin,luteolin, nepetin, nepitrin, nobiletin, orientin, oroxindin, oroxylin A,rhoifolin, scutellarein, scutellarin, tangeritin, techtochrysin, tetuin,tricin, veronicastroside, vitexin, wogonin Flavonols3-hydroxy-2-phenylchromen-4-one 3-hydroxyflavone, azaleatin, fisetin,galangin, gossypetin, kaempferide, kaempferol, isorhamnetin, morin,myricetin, natsudaidain, pachypodol, quercetin, rhamnazin, rhamnetin,sophorin Flavanones 2,3-dihydro-2-phenylchromen-4-one butin,eriodictyol, hesperetin, hesperidin, homoeriodictyol, isosakuranetin,naringenin, naringin, pinocembrin, poncirin, sakuranetin, sakuranin,sterubin Flavanonols 3-hydroxy-2,3-dihydro-2- aromadedrin, taxifolinphenylchromen-4-one Isoflavones 3-phenylchromen-4-one alpinumisoflavone,anagyroidisoflavone A and B, calycosin, daidzein, daidzin, derrubone,di-O- methylalpinumisoflavone, formononetin, genistein, genistin,glycitein, ipriflavone, irigenin, iridin, irilone, 4′-methyl-alpinumisoflavone, 5-O- methylgenistein, luteone, ononin, orobol,pratensein, prunetin, pseudobaptigenin, psi-tectorigenin, puerarin,retusin, tectoridin, tectorigenin, wighteone Isoflavenes 3-phenylchromanlonchocarpane, laxiflorane Neoflavones 4-phenylcoumarine calophyllolideNeoflavenes 4-phenylchromen dalbergichromene Flavan-3-ols2-phenyl-3,4-dihydro-2H-chromen- arthromerin A, arthromerin B, 3-olfisetinidol, guibourtinidol, meciadanol (3-O-methylcatechin), mesquitol,robinetinidol, thearubigin. Catechins (2R,3S)-2-(3,4-dihydroxyphenyl)-(+)-catechin (2R-3S), (−)-catechin 3,4-dihydro-2H-chromene-3,5,7-triol(2S-3R), (−)-Epicatechin (2R-3R), (+)-epicatechin (2S-3S) Flavan-4-ols2-phenylchroman-4-ol apiforol, luteoforol Flavan-3,4-2-phenyl-3,4-dihydro-2H-chromene- leucocyanidin, leucodelphinidin, diols3,4-diol leucomalvidin, leucopelargonidin, leucopeonidin,leucorobinetinidin, melacacidin Anthocyanidins 2-phenylchromenyliumantirrhinin, apigeninidin, aurantinidin, capensinidin, chrysanthenin,columnidin, commelinin, cyanidin, 6- hydroxycyanidin, cyanidin-3-(di-p-coumarylglucoside)-5-glucoside, cyanosalvianin, delphinidin,diosmetinidin, europinidin, fisetinidin, gesneridin, guibourtinidin,hirsutidin, luteolinidin, malvidin, 5-desoxy- malvidin, malvin,myrtillin, oenin, peonidin, 5-desoxy-peonidin, pelargonidin, petunidin,primulin, protocyanin, protodelphin, pulchellidin, pulchellidin 3-glucoside, pulchellidin 3- rhamnoside, robinetinidin, rosinidin,tricetinidin, tulipanin, violdelphin Hydrolyzable gallic acid or ellagicacid castalagin, castalin, casuarictin, tannins casuariin, casuarinin,corilagin, cornusiin E, grandinin, galloyl glucose, digalloyl glucose,trigalloyl glucose, tetragalloyl glucose, pentagalloyl glucose,hexagalloyl glucose, heptagalloyl glucose, octagalloyl glucose,pedunculagin, punicacortein C, punigluconin, punicalagin, punicalaginalpha, punicalin, 2-O-galloyl-punicalin, stachyurin, strictinin, tannicacid, tellimagrandin II Condensed polymer chains of flavonoid unitsproanthocyanidin, prodelphinidin, tannins profisetinidin,proguibourtinidin, prorobinetidin

The amount of a flavonoid included in a fluid composition disclosed inthe present specification is an amount effective to reduce or preventdegradation of a matrix polymer disclosed in the present specification.As such, the amount of a flavonoid included in a fluid compositiondisclosed in the present specification is between about 0.1% to about10% by weight of the total composition. In addition, a fluid compositiondisclosed in the present specification may comprise a single flavonoid,or a plurality of flavonoid. Further, a fluid composition disclosed inthe present specification comprises a flavonoid that is pharmaceuticallyacceptable.

Thus in an embodiment, a fluid composition comprises a pharmaceuticallyacceptable flavonoid that can reduce or prevent degradation of a matrixpolymer. In aspects of this embodiment, a fluid composition comprises apharmaceutically acceptable C-methylated flavonoid, a pharmaceuticallyacceptable O-methylated flavonoid, a pharmaceutically acceptableisoflavonoid, a pharmaceutically acceptable neoflavonoid, apharmaceutically acceptable flavonolignan, a pharmaceutically acceptablefuranoflavonoid, a pharmaceutically acceptable pyranoflavonoid, apharmaceutically acceptable methylenedioxyflavonoid, a pharmaceuticallyacceptable prenylated flavonoid, a pharmaceutically acceptable aurone, apharmaceutically acceptable flavone, a pharmaceutically acceptableflavonol, a pharmaceutically acceptable flavanone, a pharmaceuticallyacceptable flavanonol, a pharmaceutically acceptable flavan-3-ol, apharmaceutically acceptable flavan-4-ol, a pharmaceutically acceptableleucoanthocyanidin, a pharmaceutically acceptable anthocyanidin, and apharmaceutically acceptable tannin.

In another embodiment, a fluid composition comprises a single flavonoidthat can reduce or prevent degradation of a matrix polymer. In yetanother embodiment, a fluid composition comprises a plurality offlavonoids, each of which can reduce or prevent the degradation of amatrix polymer. In aspects of this embodiment, a fluid compositioncomprises one or more flavonoids, two or more flavonoids, three or moreflavonoids, four or more flavonoids, or five or more flavonoids. Inother aspects of this embodiment, a fluid composition comprises one tofive flavonoids, two to five flavonoids, three to five flavonoids, twoto four flavonoids, two to five flavonoids, or three to five flavonoids.

In yet another embodiment, a fluid composition comprises a flavonoid inan amount sufficient to reduce or prevent degradation of a matrixpolymer. In aspects of this embodiment, a fluid composition comprises aflavonoid in an amount of, e.g., about 0.1% (w/v) of the composition,about 0.2% (w/v) of the composition, about 0.3% (w/v) of thecomposition, about 0.4% (w/v) of the composition, about 0.5% (w/v) ofthe composition, about 0.6% (w/v) of the composition, about 0.7% (w/v)of the composition, about 0.8% (w/v) of the composition, about 0.9%(w/v) of the composition, about 1.0% (w/v) of the composition, about2.0% (w/v) of the composition, about 3.0% (w/v) of the composition,about 4.0% (w/v) of the composition, about 5.0% (w/v) of thecomposition, about 6.0% (w/v) of the composition, about 7.0% (w/v) ofthe composition, about 8.0% (w/v) of the composition, about 9.0% (w/v)of the composition, or about 10% (w/v) of the composition. In otheraspects, a fluid composition comprises a flavonoid in an amount of,e.g., at least 0.1% (w/v) of the composition, at least 0.2% (w/v) of thecomposition, at least 0.3% (w/v) of the composition, at least 0.4% (w/v)of the composition, at least 0.5% (w/v) of the composition, at least0.6% (w/v) of the composition, at least 0.7% (w/v) of the composition,at least 0.8% (w/v) of the composition, at least 0.9% (w/v) of thecomposition, at least 1.0% (w/v) of the composition, at least 2.0% (w/v)of the composition, at least 3.0% (w/v) of the composition, at least4.0% (w/v) of the composition, at least 5.0% (w/v) of the composition,at least 6.0% (w/v) of the composition, at least 7.0% (w/v) of thecomposition, at least 8.0% (w/v) of the composition, at least 9.0% (w/v)of the composition, or at least 10% (w/v) of the composition. In yetother aspects, a fluid composition comprises a flavonoid in an amountof, e.g., at most 0.1% (w/v) of the composition, at most 0.2% (w/v) ofthe composition, at most 0.3% (w/v) of the composition, at most 0.4%(w/v) of the composition, at most 0.5% (w/v) of the composition, at most0.6% (w/v) of the composition, at most 0.7% (w/v) of the composition, atmost 0.8% (w/v) of the composition, at most 0.9% (w/v) of thecomposition, at most 1.0% (w/v) of the composition, at most 2.0% (w/v)of the composition, at most 3.0% (w/v) of the composition, at most 4.0%(w/v) of the composition, at most 5.0% (w/v) of the composition, at most6.0% (w/v) of the composition, at most 7.0% (w/v) of the composition, atmost 8.0% (w/v) of the composition, at most 9.0% (w/v) of thecomposition, or at most 10% (w/v) of the composition. In still otheraspects, a fluid composition comprises a flavonoid in an amount of,e.g., about 0.1% (w/v) to about 1.0% (w/v) of the composition, about0.1% (w/v) to about 2.0% (w/v) of the composition, about 0.1% (w/v) toabout 3.0% (w/v) of the composition, about 0.1% (w/v) to about 4.0%(w/v) of the composition, about 0.1% (w/v) to about 5.0% (w/v) of thecomposition, about 0.2% (w/v) to about 0.9% (w/v) of the composition,about 0.2% (w/v) to about 1.0% (w/v) of the composition, about 0.2%(w/v) to about 2.0% (w/v) of the composition, about 0.5% (w/v) to about1.0% (w/v) of the composition, or about 0.5% (w/v) to about 2.0% (w/v)of the composition.

Aspects of the present specification provide, in part, a fluidcomposition that can optionally comprise or not comprise another activeingredient. As used herein, the term “active ingredient” includes but isnot limited to a drug. A drug can generally be defined as a chemicalsubstance used in the treatment, cure, prevention, or diagnosis ofdisease or used to otherwise enhance physical or mental well-being.

Aspects of the present specification provide, in part, a fluidcomposition that can optionally comprise or not comprise an anti-itchagent. The amount of an anti-itch agent included in a fluid compositiondisclosed in the present specification is an amount effective tomitigate an itch response experienced by an individual uponadministration of the composition. As such, the amount of an anti-itchagent included in a fluid composition disclosed in the presentspecification is between about 0.1% to about 5% by weight of the totalcomposition. Non-limiting examples of anti-itch agents include methylsulphonyl methane, sodium bicarbonate, calamine, allantoin, kaolin,peppermint, tea tree oil and combinations thereof.

Thus in an embodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-itch agent. In aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-itch agent where the amount ofanti-itch agent present is about 0.1% (w/v) to about 5% (w/v) of thetotal composition, about 0.1% (w/v) to about 1% (w/v) of the totalcomposition, or about 0.1% (w/v) to about 0.5% (w/v) of the totalcomposition. In other aspects of this embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, and an anti-itchagent where the amount of anti-itch agent present is about 0.3%. Inanother aspect of this embodiment, a fluid composition comprises amatrix polymer, a stabilizing component, and methyl sulphonyl methane,sodium bicarbonate, calamine, allantoin, kaolin, peppermint, tea treeoil, or combinations thereof. In aspects of this embodiment, a fluidcomposition comprises a matrix polymer, a stabilizing component, and amethyl sulphonyl methane, sodium bicarbonate, calamine, allantoin,kaolin, peppermint, tea tree oil, or combinations thereof where theamount of methyl sulphonyl methane, sodium bicarbonate, calamine,allantoin, kaolin, peppermint, or tea tree oil present is about 0.1%(w/v) to about 5% (w/v) of the total composition, about 0.1% (w/v) toabout 1% (w/v) of the total composition, or about 0.1% (w/v) to about0.5% (w/v) of the total composition. In other aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and a methyl sulphonyl methane, sodiumbicarbonate, calamine, allantoin, kaolin, peppermint, tea tree oil, orcombinations thereof where the amount of methyl sulphonyl methane,sodium bicarbonate, calamine, allantoin, kaolin, peppermint, or tea treeoil present is about 0.3%. In another embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, but not ananti-itch agent.

Aspects of the present specification provide, in part, a fluidcomposition that can optionally comprise or not comprise ananti-cellulite agent. The amount of an anti-cellulite agent included ina fluid composition disclosed in the present specification is an amounteffective to mitigate a fatty deposit experienced by an individual uponadministration of the composition. As such, the amount of ananti-cellulite agent included in a fluid composition disclosed in thepresent specification is between about 0.1% to about 5% by weight of thetotal composition. Non-limiting examples of anti-cellulite agentsinclude forskolin, xanthine compounds such as, but not limited to,caffeine, theophylline, theobromine, and aminophylline, and combinationsthereof.

Thus in an embodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-cellulite agent. In aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-cellulite agent where the amount ofanti-cellulite agent present is about 0.1% (w/v) to about 5% (w/v) ofthe total composition, about 0.1% (w/v) to about 1% (w/v) of the totalcomposition, or about 0.1% (w/v) to about 0.5% (w/v) of the totalcomposition. In other aspects of this embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, and ananti-cellulite agent where the amount of anti-cellulite agent present isabout 0.3%. In another aspect of this embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, and forskolin, axanthine compound, or combinations thereof. In aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and a forskolin, a xanthine compound, orcombinations thereof where the amount of forskolin or a xanthinecompound present is about 0.1% (w/v) to about 5% (w/v) of the totalcomposition, about 0.1% (w/v) to about 1% (w/v) of the totalcomposition, or about 0.1% (w/v) to about 0.5% (w/v) of the totalcomposition. In other aspects of this embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, and a forskolin, axanthine compound, or combinations thereof where the amount of forskolinor a xanthine compound present is about 0.3%. In another embodiment, afluid composition comprises a matrix polymer, a stabilizing component,but not an anti-cellulite agent.

Aspects of the present specification provide, in part, a fluidcomposition that can optionally comprise or not comprise ananti-scarring agent. The amount of an anti-scarring agent included in afluid composition disclosed in the present specification is an amounteffective to mitigate a scaring response experienced by an individualupon administration of the composition. As such, the amount of ananti-scarring agent included in a fluid composition disclosed in thepresent specification is between about 0.1% to about 5% by weight of thetotal composition. Non-limiting examples of anti-scarring agents includeIFN-γ, fluorouracil, poly(lactic-co-glycolic acid), methylatedpolyethylene glycol, polylactic acid, polyethylene glycol andcombinations thereof.

Thus in an embodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-scarring agent. In aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-scarring agent where the amount ofanti-scarring agent present is about 0.1% (w/v) to about 5% (w/v) of thetotal composition, about 0.1% (w/v) to about 1% (w/v) of the totalcomposition, or about 0.1% (w/v) to about 0.5% (w/v) of the totalcomposition. In other aspects of this embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, and ananti-scarring agent where the amount of anti-scarring agent present isabout 0.3%. In another aspect of this embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, and IFN-γ,fluorouracil, poly(lactic-co-glycolic acid), methylated polyethyleneglycol, polylactic acid, polyethylene glycol, or combinations thereof.In aspects of this embodiment, a fluid composition comprises a matrixpolymer, a stabilizing component, and a IFN-γ, fluorouracil,poly(lactic-co-glycolic acid), methylated polyethylene glycol,polylactic acid, polyethylene glycol, or combinations thereof where theamount of IFN-γ, fluorouracil, poly(lactic-co-glycolic acid), methylatedpolyethylene glycol, polylactic acid, or polyethylene glycol present isabout 0.1% (w/v) to about 5% (w/v) of the total composition, about 0.1%(w/v) to about 1% (w/v) of the total composition, or about 0.1% (w/v) toabout 0.5% (w/v) of the total composition. In other aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and a IFN-γ, fluorouracil,poly(lactic-co-glycolic acid), methylated polyethylene glycol,polylactic acid, polyethylene glycol, or combinations thereof where theamount of IFN-γ, fluorouracil, poly(lactic-co-glycolic acid), methylatedpolyethylene glycol, polylactic acid, or polyethylene glycol present isabout 0.3%. In another embodiment, a fluid composition comprises amatrix polymer, a stabilizing component, but not an anti-scarring agent.

Aspects of the present specification provide, in part, a fluidcomposition that can optionally comprise or not comprise ananti-inflammatory agent. The amount of an anti-inflammatory agentincluded in a fluid composition disclosed in the present specificationis an amount effective to mitigate an inflammatory response experiencedby an individual upon administration of the composition. As such, theamount of an anti-inflammatory agent included in a fluid compositiondisclosed in the present specification is between about 0.1% to about 5%by weight of the total composition. Non-limiting examples ofanti-inflammatory agents include dexamethasone, prednisolone,corticosterone, budesonide, estrogen, sulfasalazine, mesalamine andcombinations thereof.

Thus in an embodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-inflammatory agent. In aspects ofthis embodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anti-inflammatory agent where the amountof anti-inflammatory agent present is about 0.1% (w/v) to about 5% (w/v)of the total composition, about 0.1% (w/v) to about 1% (w/v) of thetotal composition, or about 0.1% (w/v) to about 0.5% (w/v) of the totalcomposition. In other aspects of this embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, and ananti-inflammatory agent where the amount of anti-inflammatory agentpresent is about 0.3%. In another aspect of this embodiment, a fluidcomposition comprises a matrix polymer, a stabilizing component, anddexamethasone, prednisolone, corticosterone, budesonide, estrogen,sulfasalazine, mesalamine, or combinations thereof. In aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and a dexamethasone, prednisolone,corticosterone, budesonide, estrogen, sulfasalazine, mesalamine, orcombinations thereof where the amount of dexamethasone, prednisolone,corticosterone, budesonide, estrogen, sulfasalazine, or mesalaminepresent is about 0.1% (w/v) to about 5% (w/v) of the total composition,about 0.1% (w/v) to about 1% (w/v) of the total composition, or about0.1% (w/v) to about 0.5% (w/v) of the total composition. In otheraspects of this embodiment, a fluid composition comprises a matrixpolymer, a stabilizing component, and a dexamethasone, prednisolone,corticosterone, budesonide, estrogen, sulfasalazine, mesalamine, orcombinations thereof where the amount of dexamethasone, prednisolone,corticosterone, budesonide, estrogen, sulfasalazine, or mesalaminepresent is about 0.3%. In another embodiment, a fluid compositioncomprises a matrix polymer, a stabilizing component, but not ananesthetic agent.

Aspects of the present specification provide, in part, a fluidcomposition that can optionally comprise or not comprise an anestheticagent. An anesthetic agent is preferably a local anesthetic agent, i.e.,an anesthetic agent that causes a reversible local anesthesia and a lossof nociception, such as, e.g., aminoamide local anesthetics andaminoester local anesthetics. The amount of an anesthetic agent includedin a fluid composition disclosed in the present specification is anamount effective to mitigate pain experienced by an individual uponadministration of the composition. As such, the amount of an anestheticagent included in a fluid composition disclosed in the presentspecification is between about 0.1% to about 5% by weight of the totalcomposition. Non-limiting examples of anesthetic agents includelidocaine, ambucaine, amolanone, amylocaine, benoxinate, benzocaine,betoxycaine, biphenamine, bupivacaine, butacaine, butamben,butanilicaine, butethamine, butoxycaine, carticaine, chloroprocaine,cocaethylene, cocaine, cyclomethycaine, dibucaine, dimethisoquin,dimethocaine, diperodon, dicyclonine, ecgonidine, ecgonine, ethylchloride, etidocaine, beta-eucaine, euprocin, fenalcomine, formocaine,hexylcaine, hydroxytetracaine, isobutyl p-aminobenzoate, leucinocainemesylate, levoxadrol, lidocaine, mepivacaine, meprylcaine,metabutoxycaine, methyl chloride, myrtecaine, naepaine, octacaine,orthocaine, oxethazaine, parethoxycaine, phenacaine, phenol,piperocaine, piridocaine, polidocanol, pramoxine, prilocaine, procaine,propanocaine, proparacaine, propipocaine, propoxycaine, pseudococaine,pyrrocaine, ropivacaine, salicyl alcohol, tetracaine, tolycaine,trimecaine, zolamine, combinations thereof, and salts thereof.Non-limiting examples of aminoester local anesthetics include procaine,chloroprocaine, cocaine, cyclomethycaine, dimethocaine (larocaine),propoxycaine, procaine (novocaine), proparacaine, tetracaine(amethocaine). Non-limiting examples of aminoamide local anestheticsinclude articaine, bupivacaine, cinchocaine (dibucaine), etidocaine,levobupivacaine, lidocaine (lignocaine), mepivacaine, piperocaine,prilocaine, ropivacaine, and trimecaine. A non-limiting example of acombination local anesthetic is lidocaine/prilocaine (EMLA).

Thus in an embodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an anesthetic agent and salts thereof. Inaspects of this embodiment, a fluid composition comprises a matrixpolymer, a stabilizing component, and an aminoamide local anesthetic andsalts thereof or an aminoester local anesthetic and salts thereof. Inother aspects of this embodiment, a fluid composition comprises a matrixpolymer, a stabilizing component, and procaine, chloroprocaine, cocaine,cyclomethycaine, dimethocaine, propoxycaine, procaine, proparacaine,tetracaine, or salts thereof. In yet other aspects of this embodiment, afluid composition comprises a matrix polymer, a stabilizing component,and articaine, bupivacaine, cinchocaine, etidocaine, levobupivacaine,lidocaine, mepivacaine, piperocaine, prilocaine, ropivacaine,trimecaine, or salts thereof. In still other aspects of this embodiment,a fluid composition comprises a matrix polymer, a stabilizing component,and lidocaine/prilocaine combination.

In other aspects of this embodiment, a fluid composition comprises amatrix polymer, a stabilizing component, and an anesthetic agent wherethe amount of anesthetic agent present is about 0.1% (w/v) to about 5%(w/v) of the total composition, about 0.1% (w/v) to about 1% (w/v) ofthe total composition, or about 0.1% (w/v) to about 0.5% (w/v) of thetotal composition. In other aspects of this embodiment, a fluidcomposition comprises a matrix polymer, a stabilizing component, and ananesthetic agent where the amount of anesthetic agent present is about0.3%. In other aspects of this embodiment, a fluid composition comprisesa matrix polymer, a stabilizing component, and an aminoamide localanesthetic and salts thereof or an aminoester local anesthetic and saltsthereof where the amount of the local anesthetic present is about 0.1%(w/v) to about 5% (w/v) of the total composition, about 0.1% (w/v) toabout 1% (w/v) of the total composition, or about 0.1% (w/v) to about0.5% (w/v) of the total composition. In other aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and an aminoamide local anesthetic and saltsthereof or an aminoester local anesthetic and salts thereof where theamount of the local anesthetic agent present is about 0.3%.

In another aspect of this embodiment, a fluid composition comprises amatrix polymer, a stabilizing component, and lidocaine or a lidocainesalt. In aspects of this embodiment, a fluid composition comprises amatrix polymer, a stabilizing component, and a lidocaine or a lidocainesalt where the amount of lidocaine or a lidocaine salt present is about0.1% (w/v) to about 5% (w/v) of the total composition, about 0.1% (w/v)to about 1% (w/v) of the total composition, or about 0.1% (w/v) to about0.5% (w/v) of the total composition. In other aspects of thisembodiment, a fluid composition comprises a matrix polymer, astabilizing component, and a lidocaine or a lidocaine salt where theamount of lidocaine or a lidocaine salt present is about 0.3%.

In another embodiment, a fluid composition comprises a matrix polymer, astabilizing component, but not an anesthetic agent.

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification exhibiting a complexmodulus, an elastic modulus, a viscous modulus and a tan δ. Matrixpolymers disclosed in the present specification are viscoelastic in thatthe composition has an elastic component (solid-like such as, e.g.,crosslinked matrix polymer) and a viscous component (liquid-like suchas, e.g., uncrosslinked matrix polymer) when a force is applied (stress,deformation). The rheological attribute that described this property isthe complex modulus (G*), which defines a fluid compositions totalresistance to deformation. The complex modulus can be defined as the sumof the elastic modulus (G′) and the viscous modulus (G″). Falcone, etal., Temporary Polysaccharide Dermal Fillers: A Model for PersistenceBased on Physical Properties, Dermatol Surg. 35(8): 1238-1243 (2009);Tezel, supra, 2008; Kablik, supra, 2009; Beasley, supra, 2009; each ofwhich is hereby incorporated by reference in its entirety. Elasticmodulus characterizes the firmness of a composition and is also known asthe storage modulus because it describes the storage of energy from themotion of the composition. The elastic modulus describes the interactionbetween elasticity and strength (G′=stress/strain) and, as such,provides a quantitative measurement of a composition's hardness orsoftness. Although depending on the speed at which the force is applied,a stiffer composition will have a higher elastic modulus and it willtake a greater force to deform the material a given distance, such as,e.g., an injection.

Viscous modulus is also known as the loss modulus because it describesthe energy that is lost as viscous dissipation. Tan δ is the ratio ofthe viscous modulus and the elastic modulus, tan δ=G′/G″. Falcone,supra, 2009. For tan δ values disclosed in the present specification, atan δ is obtained from the dynamic modulus at a frequency of 0.628rad/s. A lower tan δ corresponds to a stiffer, harder, or more elasticcomposition.

Thus, in an embodiment, a fluid composition exhibits a complex modulus.In aspects of this embodiment, a fluid composition exhibits a complexmodulus of, e.g., about 25 Pa, about 50 Pa, about 75 Pa, about 100 Pa,about 125 Pa, about 150 Pa, about 175 Pa, about 200 Pa, about 250 Pa,about 300 Pa, about 350 Pa, about 400 Pa, about 450 Pa, about 500 Pa,about 550 Pa, about 600 Pa, about 650 Pa, about 700 Pa, about 750 Pa, orabout 800 Pa. In other aspects of this embodiment, a fluid compositionexhibits a complex modulus of, e.g., at most 25 Pa, at most 50 Pa, atmost 75 Pa, at most 100 Pa, at most 125 Pa, at most 150 Pa, at most 175Pa, at most 200 Pa, at most 250 Pa, at most 300 Pa, at most 350 Pa, atmost 400 Pa, at most 450 Pa, at most 500 Pa, at most 550 Pa, at most 600Pa, at most 650 Pa, at most 700 Pa, at most 750 Pa, or at most 800 Pa.In yet other aspects of this embodiment, a fluid composition exhibits acomplex modulus of, e.g., about 25 Pa to about 150 Pa, about 25 Pa toabout 300 Pa, about 25 Pa to about 500 Pa, about 25 Pa to about 800 Pa,about 125 Pa to about 300 Pa, about 125 Pa to about 500 Pa, or about 125Pa to about 800 Pa.

In another embodiment, a fluid composition exhibits an elastic modulus.In aspects of this embodiment, a fluid composition exhibits an elasticmodulus of, e.g., about 25 Pa, about 50 Pa, about 75 Pa, about 100 Pa,about 125 Pa, about 150 Pa, about 175 Pa, about 200 Pa, about 250 Pa,about 300 Pa, about 350 Pa, about 400 Pa, about 450 Pa, about 500 Pa,about 550 Pa, about 600 Pa, about 650 Pa, about 700 Pa, about 750 Pa, orabout 800 Pa. In other aspects of this embodiment, a fluid compositionexhibits an elastic modulus of, e.g., at most 25 Pa, at most 50 Pa, atmost 75 Pa, at most 100 Pa, at most 125 Pa, at most 150 Pa, at most 175Pa, at most 200 Pa, at most 250 Pa, at most 300 Pa, at most 350 Pa, atmost 400 Pa, at most 450 Pa, at most 500 Pa, at most 550 Pa, at most 600Pa, at most 650 Pa, at most 700 Pa, at most 750 Pa, or at most 800 Pa.In yet other aspects of this embodiment, a fluid composition exhibits anelastic modulus of, e.g., about 25 Pa to about 150 Pa, about 25 Pa toabout 300 Pa, about 25 Pa to about 500 Pa, about 25 Pa to about 800 Pa,about 125 Pa to about 300 Pa, about 125 Pa to about 500 Pa, or about 125Pa to about 800 Pa.

In another embodiment, a fluid composition exhibits a viscous modulus.In aspects of this embodiment, a fluid composition exhibits a viscousmodulus of, e.g., about 10 Pa, about 20 Pa, about 30 Pa, about 40 Pa,about 50 Pa, about 60 Pa, about 70 Pa, about 80 Pa, about 90 Pa, about100 Pa, about 110 Pa, about 120 Pa, about 130 Pa, about 140 Pa, or about150 Pa. In other aspects of this embodiment, a fluid compositionexhibits a viscous modulus of, e.g., at most 10 Pa, at most 20 Pa, atmost 30 Pa, at most 40 Pa, at most 50 Pa, at most 60 Pa, at most 70 Pa,at most 80 Pa, at most 90 Pa, at most 100 Pa, at most 110 Pa, at most120 Pa, at most 130 Pa, at most 140 Pa, or at most 150 Pa. In yet otheraspects of this embodiment, a fluid composition exhibits a viscousmodulus of, e.g., about 10 Pa to about 30 Pa, about 10 Pa to about 50Pa, about 10 Pa to about 100 Pa, about 10 Pa to about 150 Pa, or about70 Pa to about 100 Pa.

In another embodiment, a fluid composition disclosed in the presentspecification exhibiting a tan δ. In aspects of this embodiment, a fluidcomposition exhibits a tan δ of, e.g., about 0.1, about 0.2, about 0.3,about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, orabout 1.0. In other aspects of this embodiment, a fluid compositionexhibits a tan δ of, e.g., at most 0.1, at most 0.2, at most 0.3, atmost 0.4, at most 0.5, at most 0.6, at most 0.7, at most 0.8, at most0.9, or at most 1.0. In yet other aspects of this embodiment, a fluidcomposition exhibits a tan δ of, e.g., about 0.1 to about 0.3, about 0.3to about 0.5, about 0.3 to about 0.6, about 0.1 to about 0.5, or about0.1 to about 0.6.

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification exhibiting a dynamicviscosity. Viscosity is resistance of a fluid to shear or flow caused byeither shear stress or tensile stress. Viscosity describes a fluid'sinternal resistance to flow caused by intermolecular friction exertedwhen layers of fluids attempt to slide by one another and may be thoughtof as a measure of fluid friction. The less viscous the fluid, thegreater its ease of movement (fluidity).

Viscosity can be defined in two ways; dynamic viscosity (μ, although ηis sometimes used) or kinematic viscosity (v). Dynamic viscosity, alsoknown as absolute or complex viscosity, is the tangential force per unitarea required to move one horizontal plane with respect to the other atunit velocity when maintained a unit distance apart by the fluid. The SIphysical unit of dynamic viscosity is the Pascal-second (Pa·s), which isidentical to N·m-2·s. Dynamic viscosity can be expressed as τ=μ dvx/dz,where τ=shearing stress, μ=dynamic viscosity, and dvx/dz is the velocitygradient over time. For example, if a fluid with a viscosity of one Pa·sis placed between two plates, and one plate is pushed sideways with ashear stress of one Pascal, it moves a distance equal to the thicknessof the layer between the plates in one second. Dynamic viscositysymbolize by is also used, is measured with various types of rheometers,devices used to measure the way in which a liquid, suspension or slurryflows in response to applied forces.

Kinematic viscosity (v) is the ratio of dynamic viscosity to density, aquantity in which no force is involved and is defined as follows: v=μ/ρ,where μ is the dynamic viscosity ρ is density with the SI unit of kg/m³.Kinematic viscosity is usually measured by a glass capillary viscometeras has an SI unit of m²/s.

The viscosity of a fluid is highly temperature dependent and for eitherdynamic or kinematic viscosity to be meaningful, the referencetemperature must be quoted. For the viscosity values disclosed in thepresent specification, a dynamic viscosity is measured at 1 Pa with acone/plane geometry 2°/40 cm and a temperature of 20° C. Examples of thedynamic viscosity of various fluids at 20° C. is as follows: water isabout 1.0×10⁻³ Pa·s, blood is about 3-4×10⁻³ Pa·s, vegetable oil isabout 60-85×10⁻³ Pa·s, motor oil SE 30 is about 0.2 Pa·s, glycerin isabout 1.4 Pa·s, maple syrup is about 2-3 Pa·s, honey is about 10 Pa·s,chocolate syrup is about 10-25 Pa·s, peanut butter is about 150-250Pa·s, lard is about 1,000 Pa·s, vegetable shortening is about 1,200Pa·s, and tar is about 30,000 Pa·s.

Thus, in an embodiment, a fluid composition comprising a matrix polymerand a stabilizing component exhibits a dynamic viscosity. In aspects ofthis embodiment, a fluid composition comprising a matrix polymer and astabilizing component exhibits a dynamic viscosity of, e.g., about 10Pa·s, about 20 Pa·s, about 30 Pa·s, about 40 Pa·s, about 50 Pa·s, about60 Pa·s, about 70 Pa·s, about 80 Pa·s, about 90 Pa·s, about 100 Pa·s,about 125 Pa·s, about 150 Pa·s, about 175 Pa·s, about 200 Pa·s, about225 Pa·s, about 250 Pa·s, about 275 Pa·s, about 300 Pa·s, about 400Pa·s, about 500 Pa·s, about 600 Pa·s, about 700 Pa·s, about 750 Pa·s,about 800 Pa·s, about 900 Pa·s, about 1,000 Pa·s, about 1,100 Pa·s, orabout 1,200 Pa·s. In other aspects of this embodiment, a fluidcomposition comprising a matrix polymer and a stabilizing componentexhibits a dynamic viscosity of, e.g., at most 10 Pa·s, at most 20 Pa·s,at most 30 Pa·s, at most 40 Pa·s, at most 50 Pa·s, at most 60 Pa·s, atmost 70 Pa·s, at most 80 Pa·s, at most 90 Pa·s, at most 100 Pa·s, atmost 125 Pa·s, at most 150 Pa·s, at most 175 Pa·s, at most 200 Pa·s, atmost 225 Pa·s, at most 250 Pa·s, at most 275 Pa·s, at most 300 Pa·s, atmost 400 Pa·s, at most 500 Pa·s, at most 600 Pa·s, at most 700 Pa·s, atmost 750 Pa·s, at most 800 Pa·s, at most 900 Pa·s, or at most 1000 Pa·s.In yet other aspects of this embodiment, a fluid composition comprisinga matrix polymer and a stabilizing component exhibits a dynamicviscosity of, e.g., about 10 Pa·s to about 100 Pa·s, about 10 Pa·s toabout 150 Pa·s, about 10 Pa·s to about 250 Pa·s, about 50 Pa·s to about100 Pa·s, about 50 Pa·s to about 150 Pa·s, about 50 Pa·s to about 250Pa·s, about 100 Pa·s to about 500 Pa·s, about 100 Pa·s to about 750Pa·s, about 100 Pa·s to about 1,000 Pa·s, about 100 Pa·s to about 1,200Pa·s, about 300 Pa·s to about 500 Pa·s, about 300 Pa·s to about 750Pa·s, about 300 Pa·s to about 1,000 Pa·s, or about 300 Pa·s to about1,200 Pa·s.

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification that is injectable.As used herein, the term “injectable” refers to a fluid compositiondisclosed in the present specification having the properties necessaryto administer the composition into a dermal region of an individualusing an injection device with a fine needle. As used herein, the term“fine needle” refers to a needle that is 27 gauge or smaller.Injectability of a fluid composition disclosed in the presentspecification can be accomplished by sizing the fluid composition, asdiscussed below.

Thus, in an embodiment, a fluid composition comprising a matrix polymerand a stabilizing component, wherein the composition is injectable. Inaspect of this embodiment, a fluid composition comprising a matrixpolymer and a stabilizing component is injectable through a fine needle.In other aspects of this embodiment, a fluid composition comprising amatrix polymer and a stabilizing component is injectable through aneedle of, e.g., about 27 gauge, about 30 gauge, or about 32 gauge. Inyet other aspects of this embodiment, a fluid composition comprising amatrix polymer and a stabilizing component is injectable through aneedle of, e.g., 27 gauge or smaller, 30 gauge or smaller, or 32 gaugeor smaller. In still other aspects of this embodiment, a fluidcomposition comprising a matrix polymer and a stabilizing component isinjectable through a needle of, e.g., about 27 gauge to about 32 gauge.

In other aspects of this embodiment, a fluid composition comprising acrosslinked matrix polymer where the mean particle size of thecrosslinked matrix polymer is, e.g., about 200 μm, about 250 μm, about300 μm, about 350 μm, about 400 μm, about 450 μm, about 500 μm, about550 μm, about 600 μm, about 650 μm, about 700 μm, about 750 μm, or about800 μm. In yet other aspects of this embodiment, a fluid compositioncomprising a crosslinked matrix polymer where the mean particle size ofthe crosslinked matrix polymer is, e.g., at most 200 μm, at most 250 μm,at most 300 μm, at most 350 μm, at most 400 μm, at most 450 μm, at most500 μm, at most 550 μm, at most 600 μm, at most 650 μm, at most 700 μm,at most 750 μm, or at most 800 μm. In still other aspects of thisembodiment, a fluid composition comprising a crosslinked matrix polymerwhere the mean particle size of the crosslinked matrix polymer is, e.g.,about 200 μm to about 300 μm, about 300 μm to about 400 μm, about 400 μmto about 500 μm, about 500 μm to about 600 μm, about 600 μm to about 700μm, about 700 μm to about 800 μm, about 200 μm to about 400 μm, about200 μm to about 500 μm, about 200 μm to about 600 μm, about 200 μm toabout 700 μm, about 200 μm to about 800 μm, about 300 μm to about 500μm, about 300 μm to about 600 μm, about 300 μm to about 700 μm, or about300 μm to about 800 μm.

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification exhibiting aphysiologically-acceptable osmolarity. As used herein, the term “aphysiologically-acceptable osmolarity” refers to an osmolarity in accordwith, or characteristic of, the normal functioning of a living organism.As such, administration of a fluid composition disclosed in the presentcomposition exhibits an osmolarity that has substantially no long termor permanent detrimental effect when administered to mammal. Osmolarityrefers to the concentration of osmotically active solutes in solution.Osmolarity is expressed in terms of osmoles of osmotically active soluteper liter of solvent (Osmol/L or Osm/L). Osmolarity is distinct frommolarity because it measures moles of osmotically active soluteparticles rather than moles of solute. The distinction arises becausesome compounds can dissociate in solution, whereas others cannot. Theosmolarity of a solution can be calculated from the followingexpression: Osmol/L=Σφ_(i)η_(i)C_(i), where φ is the osmoticcoefficient, which accounts for the degree of non-ideality of thesolution; η is the number of particles (e.g. ions) into which a moleculedissociates; and C is the molar concentration of the solute; and i isthe index representing the identity of a particular solute. Theosmolarity of a composition disclosed in the present specification canbe measured using a conventional method that measures solutions.

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification exhibiting aphysiologically-acceptable osmolality. As used herein, the term “aphysiologically-acceptable osmolality” refers to an osmolality in accordwith, or characteristic of, the normal functioning of a living organism.As such, administration of a fluid composition disclosed in the presentcomposition exhibits an osmolality that has substantially no long termor permanent detrimental effect when administered to mammal. Osmolalityrefers to the concentration of osmotically active solutes per kilo ofwater in the body and is expressed in terms of osmoles of osmoticallyactive solute per kilogram of solvent (Osmol/kg or Osm/kg) and is equalto the sum of the molalities of all the solutes present in thatsolution. The osmolality of a solution can be measured using anosmometer. The most commonly used instrument in modern laboratories is afreezing point depression osmometer. This instruments measure the changein freezing point that occurs in a solution with increasing osmolality(freezing point depression osmometer) or the change in vapor pressurethat occurs in a solution with increasing osmolality (vapor pressuredepression osmometer).

Thus, in an embodiment, a fluid composition comprising a matrix polymerand a stabilizing component exhibit a physiologically-acceptableosmolarity. In aspects of this embodiment, a fluid compositioncomprising a matrix polymer and a stabilizing component exhibit anosmolarity of, e.g., about 100 mOsm/L, about 150 mOsm/L, about 200mOsm/L, about 250 mOsm/L, about 300 mOsm/L, about 350 mOsm/L, about 400mOsm/L, about 450 mOsm/L, or about 500 mOsm/L. In other aspects of thisembodiment, a fluid composition comprising a matrix polymer and astabilizing component exhibit an osmolarity of, e.g., at least 100mOsm/L, at least 150 mOsm/L, at least 200 mOsm/L, at least 250 mOsm/L,at least 300 mOsm/L, at least 350 mOsm/L, at least 400 mOsm/L, at least450 mOsm/L, or at least 500 mOsm/L. In yet other aspects of thisembodiment, a fluid composition comprising a matrix polymer and astabilizing component exhibit an osmolarity of, e.g., at most 100mOsm/L, at most 150 mOsm/L, at most 200 mOsm/L, at most 250 mOsm/L, atmost 300 mOsm/L, at most 350 mOsm/L, at most 400 mOsm/L, at most 450mOsm/L, or at most 500 mOsm/L. In still other aspects of thisembodiment, a fluid composition comprising a matrix polymer and astabilizing component exhibit an osmolarity of, e.g., about 100 mOsm/Lto about 500 mOsm/L, about 200 mOsm/L to about 500 mOsm/L, about 200mOsm/L to about 400 mOsm/L, about 300 mOsm/L to about 400 mOsm/L, about270 mOsm/L to about 390 mOsm/L, about 225 mOsm/L to about 350 mOsm/L,about 250 mOsm/L to about 325 mOsm/L, about 275 mOsm/L to about 300mOsm/L, or about 285 mOsm/L to about 290 mOsm/L.

In another embodiment, a fluid composition comprising a matrix polymerand a stabilizing component exhibit a physiologically-acceptableosmolality. In aspects of this embodiment, a fluid compositioncomprising a matrix polymer and a stabilizing component exhibit anosmolality of, e.g., about 100 mOsm/kg, about 150 mOsm/kg, about 200mOsm/kg, about 250 mOsm/kg, about 300 mOsm/kg, about 350 mOsm/kg, about400 mOsm/kg, about 450 mOsm/kg, or about 500 mOsm/kg. In other aspectsof this embodiment, a fluid composition comprising a matrix polymer anda stabilizing component exhibit an osmolality of, e.g., at least 100mOsm/kg, at least 150 mOsm/kg, at least 200 mOsm/kg, at least 250mOsm/kg, at least 300 mOsm/kg, at least 350 mOsm/kg, at least 400mOsm/kg, at least 450 mOsm/kg, or at least 500 mOsm/kg. In yet otheraspects of this embodiment, a fluid composition comprising a matrixpolymer and a stabilizing component exhibit an osmolality of, e.g., atmost 100 mOsm/kg, at most 150 mOsm/kg, at most 200 mOsm/kg, at most 250mOsm/kg, at most 300 mOsm/kg, at most 350 mOsm/kg, at most 400 mOsm/kg,at most 450 mOsm/kg, or at most 500 mOsm/kg. In still other aspects ofthis embodiment, a fluid composition comprising a matrix polymer and astabilizing component exhibit an osmolality of, e.g., about 100 mOsm/kgto about 500 mOsm/kg, about 200 mOsm/kg to about 500 mOsm/kg, about 200mOsm/kg to about 400 mOsm/kg, about 300 mOsm/kg to about 400 mOsm/kg,about 270 mOsm/kg to about 390 mOsm/kg, about 225 mOsm/kg to about 350mOsm/kg, about 250 mOsm/kg to about 325 mOsm/kg, about 275 mOsm/kg toabout 300 mOsm/kg, or about 285 mOsm/kg to about 290 mOsm/kg.

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification that is apharmaceutical composition. As used herein, the term “pharmaceuticalcomposition” is synonymous with “pharmaceutically-acceptablecomposition” and refers to a therapeutically effective concentration ofan active ingredient, such as, e.g., any of the matrix polymersdisclosed in the present specification. A pharmaceutical compositioncomprising a matrix polymer active ingredient is useful for medical andveterinary applications. A pharmaceutical composition may beadministered to a patient alone, or in combination with othersupplementary active ingredients, agents, drugs or hormones.

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification that is apharmaceutical composition comprising a pharmacologically acceptableexcipient. As used herein, the term “pharmacologically acceptableexcipient” is synonymous with “pharmacological excipient” or “excipient”and refers to any excipient that has substantially no long term orpermanent detrimental effect when administered to mammal and encompassescompounds such as, e.g., stabilizing agent, a bulking agent, acryo-protectant, a lyo-protectant, an additive, a vehicle, a carrier, adiluent, or an auxiliary. An excipient generally is mixed with an activeingredient, or permitted to dilute or enclose the active ingredient andcan be a solid, semi-solid, or liquid agent. It is also envisioned thata pharmaceutical composition comprising a matrix polymer activeingredient can include one or more pharmaceutically acceptableexcipients that facilitate processing of an active ingredient intopharmaceutically acceptable compositions. Insofar as anypharmacologically acceptable excipient is not incompatible with thematrix polymer active ingredient, its use in pharmaceutically acceptablecompositions is contemplated. Non-limiting examples of pharmacologicallyacceptable excipients can be found in, e.g., Pharmaceutical Dosage Formsand Drug Delivery Systems (Howard C. Ansel et al., eds., LippincottWilliams & Wilkins Publishers, 7^(th) ed. 1999); Remington: The Scienceand Practice of Pharmacy (Alfonso R. Gennaro ed., Lippincott, Williams &Wilkins, 20^(th) ed. 2000); Goodman & Gilman's The Pharmacological Basisof Therapeutics (Joel G. Hardman et al., eds., McGraw-Hill Professional,10^(th) ed. 2001); and Handbook of Pharmaceutical Excipients (Raymond C.Rowe et al., APhA Publications, 4^(th) edition 2003), each of which ishereby incorporated by reference in its entirety.

It is further envisioned that a pharmaceutical composition disclosed inthe present specification may optionally include or not include, withoutlimitation, other pharmaceutically acceptable components (orpharmaceutical components), including, without limitation, buffers,preservatives, tonicity adjusters, salts, antioxidants, osmolalityadjusting agents, emulsifying agents, wetting agents, sweetening orflavoring agents, and the like.

Pharmaceutically acceptable buffer is any buffer that can be used toprepare a pharmaceutical composition disclosed in the presentspecification, provided that the resulting preparation ispharmaceutically acceptable. Non-limiting examples of pharmaceuticallyacceptable buffers include acetate buffers, borate buffers, citratebuffers, neutral buffered salines, phosphate buffers, and phosphatebuffered salines. Any concentration of a pharmaceutically acceptablebuffer can be useful in formulating a pharmaceutical compositiondisclosed in the present specification, with the proviso that atherapeutically effective amount of the matrix polymer active ingredientis recovered using this effective concentration of buffer. Non-limitingexamples of concentrations of physiologically-acceptable buffers occurwithin the range of about 0.1 mM to about 900 mM. The pH ofpharmaceutically acceptable buffers may be adjusted, provided that theresulting preparation is pharmaceutically acceptable. It is understoodthat acids or bases can be used to adjust the pH of a pharmaceuticalcomposition as needed. Any buffered pH level can be useful informulating a pharmaceutical composition, with the proviso that atherapeutically effective amount of the matrix polymer active ingredientis recovered using this effective pH level. Non-limiting examples ofphysiologically-acceptable pH occur within the range of about pH 5.5 toabout pH 8.5.

Pharmaceutically acceptable antioxidants include, without limitation,sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylatedhydroxyanisole and butylated hydroxytoluene. Pharmaceutically acceptablepreservatives include, without limitation, benzalkonium chloride,chlorobutanol, thimerosal, phenylmercuric acetate, phenylmercuricnitrate, a stabilized oxy chloro composition, such as, e.g., PURITE®(Allergan, Inc. Irvine, Calif.) and chelants, such as, e.g., DTPA orDTPA-bisamide, calcium DTPA, and CaNaDTPA-bisamide.

Tonicity adjustors useful in a pharmaceutical composition include,without limitation, salts such as, e.g., sodium chloride and potassiumchloride; and glycerin. The pharmaceutical composition may be providedas a salt and can be formed with many acids, including but not limitedto, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic,etc. Salts tend to be more soluble in aqueous or other protonic solventsthan are the corresponding free base forms. It is understood that theseand other substances known in the art of pharmacology can be included ina pharmaceutical composition useful in the invention. Other non-limitingexamples of pharmacologically acceptable components can be found in,e.g., Ansel, supra, (1999); Gennaro, supra, (2000); Hardman, supra,(2001); and Rowe, supra, (2003), each of which is hereby incorporated byreference in its entirety.

A pharmaceutical compositions disclosed in the present specificationgenerally is administered as a pharmaceutical acceptable compositioncomprising a matrix polymer active ingredient. As used herein, the term“pharmaceutically acceptable” means any molecular entity or compositionthat does not produce an adverse, allergic or other untoward or unwantedreaction when administered to an individual.

Aspects of the present specification provide, in part, a method ofmaking a fluid composition disclosed in the present specification. In anaspect, a method for making a fluid composition, the method comprisingthe steps of: a) combining a stabilizing component with aphysiologically-acceptable buffer to make a stabilizingcomponent-buffered solution; b) combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymer,and; c) sizing the fluid composition. This method may, or may not,further comprise a step comprising titrating a stabilizingcomponent-buffered solution to obtain a desired pH after step (a); astep comprising filtering the stabilizing component-buffered solutionafter step (a); a step (b) where combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymeroccurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time; a step (b) where combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymeroccurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time and then followed by a rest for a relative long period of time;a step (b) where combining a matrix polymer with the stabilizingcomponent-buffered solution to hydrate the matrix polymer occurs bymixing the matrix polymer with the stabilizing component-bufferedsolution at a low speed for a relatively long period of time and then bymixing the matrix polymer with the stabilizing component-bufferedsolution using a cycle of alternating periods of agitation for arelatively short period of time followed by periods of rest for arelatively long period of time; a step (b) where combining a matrixpolymer with the stabilizing component-buffered solution to hydrate thematrix polymer occurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time and then by mixing the matrix polymer with the stabilizingcomponent-buffered solution using a cycle of alternating periods ofagitation for a relatively short period of time followed by periods ofrest for a relatively long period of time, and then followed by a restfor a relative long period of time; a step comprising degassing a fluidcomposition after step (b) or step (c); a step comprising filling asyringe with a fluid composition after step (c); and/or a stepcomprising sterilizing a syringe filled with a fluid composition afterstep (c).

Aspects of the present specification provide, in part, a fluidcomposition disclosed in the present specification made by a methoddisclosed in the present specification. In an aspect, a fluidcomposition comprises a matrix polymer and a stabilizing componentwherein the fluid composition is made by a method comprising the stepsof: a) combining a stabilizing component with aphysiologically-acceptable buffer to make a stabilizingcomponent-buffered solution; b) combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymer,and; c) sizing the fluid composition. This method may, or may not,further comprise a step comprising titrating a stabilizingcomponent-buffered solution to obtain a desired pH after step (a); astep comprising filtering the stabilizing component-buffered solutionafter step (a); a step (b) where combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymeroccurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time; a step (b) where combining a matrix polymer with thestabilizing component-buffered solution to hydrate the matrix polymeroccurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time and then followed by a rest for a relative long period of time;a step (b) where combining a matrix polymer with the stabilizingcomponent-buffered solution to hydrate the matrix polymer occurs bymixing the matrix polymer with the stabilizing component-bufferedsolution at a low speed for a relatively long period of time and then bymixing the matrix polymer with the stabilizing component-bufferedsolution using a cycle of alternating periods of agitation for arelatively short period of time followed by periods of rest for arelatively long period of time; a step (b) where combining a matrixpolymer with the stabilizing component-buffered solution to hydrate thematrix polymer occurs by mixing the matrix polymer with the stabilizingcomponent-buffered solution at a low speed for a relatively long periodof time and then by mixing the matrix polymer with the stabilizingcomponent-buffered solution using a cycle of alternating periods ofagitation for a relatively short period of time followed by periods ofrest for a relatively long period of time, and then followed by a restfor a relative long period of time; a step comprising degassing a fluidcomposition after step (b) or step (c); a step comprising filling asyringe with a fluid composition after step (c); and/or a stepcomprising sterilizing a syringe filled with a fluid composition afterstep (c).

Aspects of the present specification provide, in part, a method having astep of combining a stabilizing component with aphysiologically-acceptable buffer to make a stabilizingcomponent-buffered solution. A stabilizing component can be any one ofthe stabilizing components disclosed in the present specification. Asused herein, the term “a physiologically-acceptable buffer” refers to abuffer in accord with, or characteristic of, the normal functioning of aliving organism. As such, a buffer used to make a fluid compositiondisclosed in the present specification exhibits a buffering capacitythat has substantially no long term or permanent detrimental effect whenadministered to mammal. Physiologically-acceptable buffers include,without limitation, acetate buffers, borate buffers, citrate buffers,neutral buffered salines, phosphate buffers, and phosphate bufferedsalines. In addition, the physiologically-acceptable buffer is at aconcentration to achieve an effective buffering capacity. Non-limitingexamples of a concentration to a physiologically-acceptable buffer toachieve an effective buffering capacity is from between about 0.1 mM toabout 900 mM.

Thus, in an embodiment, a stabilizing component is combined with aphysiologically-acceptable buffer to make a stabilizingcomponent-buffered solution. In aspects of this embodiment, astabilizing component is combined with an acetate buffer, a boratebuffers, a citrate buffer, a neutral buffered saline, a phosphatebuffer, or a phosphate buffered saline to make a stabilizing-bufferedsolution. In other an stabilizing component is combined with sodiumchloride, sodium phosphate, or both.

In another embodiment, a physiologically-acceptable buffer is at aconcentration necessary to achieve an effective buffering capacity. Inaspects of this embodiment, a physiologically-acceptable buffer is at aconcentration of, e.g., at least 0.1 mM, at least 0.2 mM, at least 0.3mM, at least 0.4 mM, at least 0.5 mM, at least 0.6 mM, at least 0.7 mM,at least 0.8 mM, or at least 0.9 mM. In other aspects of thisembodiment, a physiologically-acceptable buffer is at a concentrationof, e.g., at least 1.0 mM, at least 2.0 mM, at least 3.0 mM, at least4.0 mM, at least 5.0 mM, at least 6.0 mM, at least 7.0 mM, at least 8.0mM, or at least 9.0 mM. In yet other aspects of this embodiment, aphysiologically-acceptable buffer is at a concentration of, e.g., atleast 10 mM, at least 20 mM, at least 30 mM, at least 40 mM, at least 50mM, at least 60 mM, at least 70 mM, at least 80 mM, or at least 90 mM.In still other aspects of this embodiment, a physiologically-acceptablebuffer is at a concentration of, e.g., at least 100 mM, at least 200 mM,at least 300 mM, at least 400 mM, at least 500 mM, at least 600 mM, atleast 700 mM, at least 800 mM, or at least 900 mM.

In further aspects of this embodiment, a physiologically-acceptablebuffer is at a concentration of, e.g., at most 0.1 mM, at most 0.2 mM,at most 0.3 mM, at most 0.4 mM, at most 0.5 mM, at most 0.6 mM, at most0.7 mM, at most 0.8 mM, or at most 0.9 mM. In still other aspects ofthis embodiment, a physiologically-acceptable buffer is at aconcentration of, e.g., at most 1.0 mM, at most 2.0 mM, at most 3.0 mM,at most 4.0 mM, at most 5.0 mM, at most 6.0 mM, at most 7.0 mM, at most8.0 mM, or at most 9.0 mM. In yet other aspects of this embodiment, aphysiologically-acceptable buffer is at a concentration of, e.g., atmost 10 mM, at most 20 mM, at most 30 mM, at most 40 mM, at most 50 mM,at most 60 mM, at most 70 mM, at most 80 mM, or at most 90 mM. In stillother aspects of this embodiment, a physiologically-acceptable buffer isat a concentration of, e.g., at most 100 mM, at most 200 mM, at most 300mM, at most 400 mM, at most 500 mM, at most 600 mM, at most 700 mM, atmost 800 mM, or at most 900 mM. In still further aspects of thisembodiment, a physiologically-acceptable buffer is at a concentrationof, e.g., about 0.1 mM to about 900 mM, 0.1 mM to about 500 mM, 0.1 mMto about 100 mM, 0.1 mM to about 90 mM, 0.1 mM to about 50 mM, 1.0 mM toabout 900 mM, 1.0 mM to about 500 mM, 1.0 mM to about 100 mM, 1.0 mM toabout 90 mM, or 1.0 mM to about 50 mM.

Aspects of the present specification provide, in part, a method havingan optional step of titrating a stabilizing component-buffered solutionto obtain a desired pH. A stabilizing component-buffered solution can betitrated to any physiologically-acceptable pH desired. As used herein,the term “a physiologically-acceptable pH” refers to a pH in accordwith, or characteristic of, the normal functioning of a living organism.As such, a pH used to make a fluid composition disclosed in the presentspecification is a pH that has substantially no long term or permanentdetrimental effect when administered to mammal. Non-limiting examples ofphysiologically-acceptable pH occur within the range of about pH 5.5 toabout pH 8.5. It is understood that acids or bases can be used to adjustthe pH of a stabilizing component-buffered solution.

Thus, in an embodiment, a stabilizing component-buffered solution istitrated to a physiologically-acceptable pH. In an aspect of thisembodiment, a stabilizing component-buffered solution is titrated to pHof, e.g., at least about pH 5.0, at least about pH 5.5, at least aboutpH 6.0, at least about pH 6.5, at least about pH 7.0 or at about pH 7.5.In another aspect of this embodiment, a stabilizing component-bufferedsolution is titrated to pH of, e.g., at most about pH 5.0, at most aboutpH 5.5, at most about pH 6.0, at most about pH 6.5, at most about pH 7.0or at most about pH 7.5. In yet another aspect of this embodiment, astabilizing component-buffered solution is titrated to pH of, e.g.,about pH 5.0 to about pH 8.0, an effective pH level is about pH 5.0 toabout pH 7.0, an effective pH level is about pH 5.0 to about pH 6.0, isabout pH 5.5 to about pH 8.0, an effective pH level is about pH 5.5 toabout pH 7.0, an effective pH level is about pH 5.5 to about pH 5.0, isabout pH 5.5 to about pH 7.5, an effective pH level is about pH 5.5 toabout pH 6.5.

Aspects of the present specification provide, in part, a method havingan optional step of filtering a stabilizing component-buffered solutionto remove particulates and impurities. For example, a stabilizingcomponent-buffered solution may, or may not, be filtered to removeparticulates and impurities from the stabilizing component-bufferedsolution. The filters used should be of a pore size that sufficientlyremoves the particulates and impurities desired to be removed from astabilizing component-buffered solution. Non-limiting examples of poresizes are of the range of 5.0 μm or less.

Thus, in an embodiment, a stabilizing component-buffered solution isfiltered to remove particulates and impurities. In an aspect of thisembodiment, a stabilizing component-buffered solution is filteredthrough a pore size sufficient to remove particulates and impurities. Inother aspects of this embodiment, a stabilizing component-bufferedsolution is filtered through a pore size of, e.g., about 0.1 μm or less,about 0.25 μm or less, about 0.5 μm or less, about 0.75 μm or less,about 1 μm or less, about 2 μm or less, about 3 μm or less, about 4 μmor less, or about 5 μm or less.

Aspects of the present specification provide, in part, a method having astep of combining a matrix polymer with a stabilizing component-bufferedsolution to hydrate the matrix polymer. A matrix polymer can be any oneof the matrix polymers disclosed in the present specification, itssalts, and/or mixtures thereof. A matrix polymer can be a partiallycrosslinked matrix polymer, a substantially uncrosslinked matrixpolymer, a matrix polymer that is essentially free of a crosslinkedmatrix polymer, or a matrix polymer that is entirely free of acrosslinked matrix polymer as disclosed in the present specification.The source of the matrix polymer can be from a bacterial source or ananimal source.

Combining a matrix polymer with a stabilizing component-bufferedsolution can be accomplished by any method with the proviso that themethod used is sufficient to hydrate the matrix polymer in a manner thatproduces a fluid composition disclosed in the present specification. Itis also understood that any method employed does not result insubstantial degradation of matrix polymer as this is inconsistent with afluid composition disclosed in the present specification. For example,the step of combining a matrix polymer with a stabilizingcomponent-buffered solution can comprises mixing a matrix polymer with astabilizing component-buffered solution at a low speed for a relativelylong period of time. Non-limiting examples of a speed used for mixing isfrom about 50 rpm to about 500 rpm. The relatively long time period foragitation is a time period sufficient to effectively combine a matrixpolymer with a stabilizing component-buffered solution to allow thematrix polymer to hydrate. Non-limiting examples of time periods formixing at a relative low speed include from about 4 hours to about 16hours. This combining step is performed at cool ambient temperature.Non-limiting examples of a cool ambient temperature include atemperature not exceeding about 25° C., such as, a temperature notexceeding about 20° C., a temperature not exceeding about 15° C., or atemperature not exceeding about 10° C. As another non-limiting example,a cool ambient temperature is a temperature from about 2° C. and about8° C.

As another non-limiting example, the step of combining a matrix polymerwith a stabilizing component-buffered solution may, or may not, involvea cycle of alternating periods of agitation for a relatively shortperiod of time followed by periods of rest for a relatively long periodof time. This agitation/rest step may be performed once in order tocombine a matrix polymer with a stabilizing component-buffered solutioncomposition, or may be performed for a plurality of times. For example,the agitation/rest step may be performed two or more times, five or moretimes, or ten or more times. Agitation of a matrix polymer with astabilizing component-buffered solution can be accomplished by anymethod sufficient to agitate the composition including, withoutlimitation, mechanical shaking, manual shaking, ultrasound, vibration,and the like, and combinations thereof. The relatively short time periodfor agitation is a time period sufficient to effectively combine amatrix polymer with a stabilizing component-buffered solution to createa fluid composition disclosed in the present specification. Non-limitingexamples of time periods for agitation include from about 1 minute toabout 15 minutes. Similarly, the relatively long time period for rest isa time period sufficient to effectively combine a matrix polymer with astabilizing component-buffered solution to create a fluid compositiondisclosed in the present specification. Non-limiting examples of timeperiods for rest include from about 15 minute to about 180 minutes. Forexample, a matrix polymer with a stabilizing component-buffered solutioncomposition can be agitated for about 1 minute and then allowed to restfor about 30 minutes. If such an agitation/rest step is used, ittypically follows the combining step comprising a low speed for arelatively long period of time described above.

After combining a matrix polymer with a stabilizing component-bufferedsolution, this composition is allowed to rest for a relative long periodof time. A relatively long time period for rest is a time periodsufficient to effectively combine a matrix polymer with a stabilizingcomponent-buffered solution to create a fluid composition disclosed inthe present specification. Non-limiting examples of time periods forresting after the combining step include from about 4 hours to about 16hours.

Thus, in an embodiment, a matrix polymer is combined with a stabilizingcomponent-buffered solution in order to hydrate the matrix polymer. Inan aspect of this embodiment, a matrix polymer is combined with astabilizing component-buffered solution in order to hydrate the matrixpolymer and achieve a smooth consistency of the composition. In otheraspects of this embodiment, a stabilizing component-buffered solution iscombined with a partially crosslinked matrix polymer, a substantiallyuncrosslinked matrix polymer, a matrix polymer that is essentially freeof a crosslinked matrix polymer, or a matrix polymer that is entirelyfree of a crosslinked matrix polymer.

In another embodiment, a matrix polymer is combined with a stabilizingcomponent-buffered solution by mixing the matrix polymer with thestabilizing component-buffered solution at a low speed for a relativelylong period of time. In aspects of this embodiment, a matrix polymer iscombined with a stabilizing component-buffered solution by mixing thematrix polymer with the stabilizing component-buffered solution at,e.g., about 50 rpm, about 100 rpm, about 150 rpm, about 200 rpm, about250 rpm, about 300 rpm, about 350 rpm, about 400 rpm, about 450 rpm, orabout 500 rpm for about 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11hours, 12 hours, 13 hours, or 14 hours. In other aspects of thisembodiment, a matrix polymer is combined with a stabilizingcomponent-buffered solution by mixing the matrix polymer with thestabilizing component-buffered solution at, e.g., about 50 rpm, about100 rpm, about 150 rpm, about 200 rpm, about 250 rpm, about 300 rpm,about 350 rpm, about 400 rpm, about 450 rpm, or about 500 rpm for about6 hours or more, 7 hours or more, 8 hours or more, 9 hours or more, 10hours or more, 11 hours or more, 12 hours or more, 13 hours or more, or14 hours or more. In yet other aspects of this embodiment, a matrixpolymer is combined with a stabilizing component-buffered solution bymixing the matrix polymer with the stabilizing component-bufferedsolution at, e.g., at most 50 rpm, at most 100 rpm, at most 150 rpm, atmost 200 rpm, at most 250 rpm, at most 300 rpm, at most 350 rpm, at most400 rpm, at most 450 rpm, or at most 500 rpm for about 6 hours or more,7 hours or more, 8 hours or more, 9 hours or more, 10 hours or more, 11hours or more, 12 hours or more, 13 hours or more, or 14 hours or more.In still other aspects of this embodiment, a matrix polymer is combinedwith a stabilizing component-buffered solution by mixing the matrixpolymer with the stabilizing component-buffered solution at about 50 rpmto about 500 rpm for about 8 hours to about 12 hours.

In another embodiment, a matrix polymer is combined with a stabilizingcomponent-buffered solution by mixing the matrix polymer with thestabilizing component-buffered solution using a cycle of alternatingperiods of agitation for a relatively short period of time followed byperiods of rest for a relatively long period of time. In aspects of thisembodiment, a matrix polymer and a stabilizing component-bufferedsolution is agitated for about 1 minute, about 5 minutes, about 10minutes, or about 15 minutes, and then allowed to rest for about 15minutes, about 30 minutes, about 45 minutes, about 60 minutes, about 75minutes, about 90 minutes, about 105 minutes, about 120 minutes, about135 minutes, about 150 minutes, about 165 minutes, or about 180 minutes.In other aspects of this embodiment, a matrix polymer and a stabilizingcomponent-buffered solution is agitated for about 1 minute or more,about 5 minutes or more, about 10 minutes or more, or about 15 minutesor more, and then allowed to rest for about 15 minutes or more, about 30minutes or more, about 45 minutes or more, about 60 minutes or more,about 75 minutes or more, about 90 minutes or more, about 105 minutes,about 120 minutes or more, about 135 minutes or more, about 150 minutesor more, about 165 minutes or more, or about 180 minutes or more. In yetother aspects of this embodiment, a matrix polymer and a stabilizingcomponent-buffered solution is agitated for at most 1 minute, at most 5minutes, at most 10 minutes, or at most 15 minutes, and then allowed torest for at most 15 minutes, at most 30 minutes, at most 45 minutes, atmost 60 minutes, at most 75 minutes, at most 90 minutes, at most 105minutes, at most 120 minutes, at most 135 minutes, at most 150 minutes,at most 165 minutes, or at most 180 minutes. In still other aspects ofthis embodiment, a matrix polymer and a stabilizing component-bufferedsolution is agitated for at most 1 minute, at most 5 minutes, at most 10minutes, or at most 15 minutes, and then allowed to rest for about 15minutes or more, about 30 minutes or more, about 45 minutes or more,about 60 minutes or more, about 75 minutes or more, about 90 minutes ormore, about 105 minutes, about 120 minutes or more, about 135 minutes ormore, about 150 minutes or more, about 165 minutes or more, or about 180minutes or more. In further aspects of this embodiment, a matrix polymerand a stabilizing component-buffered solution is agitated for about 1minute to about 15 minutes, and then allowed to rest for about 30minutes to about 60 minutes.

Aspects of the present specification provide, in part, a method having astep of sizing the fluid composition. Sizing the fluid composition isparticularly important if crosslinked matrix polymers are presentbecause this step 1) produces a composition with a particular mean sizedof gel particle, and/or 2) produces a composition with a smoothconsistency as opposed to a granular consistency. In fluid compositionscomprising a crosslinked matrix polymer, the initial crosslinkingprocess produces a large gel mass that must be sized down in order toproduce a composition that can be properly administered to anindividual, such as, e.g., by injection. Sizing of gel particles can beaccomplished by any method suitable to produce a fluid composition thatcan be properly administered into an individual. Non-limiting examplesinclude sieving and homogenization. In a sieving method, a large gelmass included in a fluid composition is broken down by passing through aseries of sieves or screens in order to size the gel particles. Thismethod produces gel particles that have a well-defined average size. Ina homogenization method, a large gel mass included in a fluidcomposition is broken down by recirculating the fluid compositionbetween a first vessel and a second vessel through a narrow aperture.The recirculation of a fluid composition may comprise passing thecomposition from a first vessel into a second vessel through an orificehaving any diameter sufficient to achieve a smooth fluid composition.Non-limiting examples of orifice diameters include about 2 mm to about10 mm. The recirculating step can be performed once or a plurality oftimes, such as, e.g., from 2 to 10 times.

Thus, in an embodiment, a fluid composition is sized. In other aspectsof this embodiment, a fluid composition is sized by sieving whereinafter sizing the mean particle size of the crosslinked matrix polymeris, e.g., about 200 μm, about 250 μm, about 300 μm, about 350 μm, about400 μm, about 450 μm, about 500 μm, about 550 μm, about 600 μm, about650 μm, about 700 μm, about 750 μm, or about 800 μm. In yet otheraspects of this embodiment, a fluid composition is sized by sievingwherein after sizing the mean particle size of the crosslinked matrixpolymer is, e.g., at most 200 μm, at most 250 μm, at most 300 μm, atmost 350 μm, at most 400 μm, at most 450 μm, at most 500 μm, at most 550μm, at most 600 μm, at most 650 μm, at most 700 μm, at most 750 μm, orat most 800 μm. In still other aspects of this embodiment, a fluidcomposition is sized by sieving wherein after sizing the mean particlesize of the crosslinked matrix polymer is, e.g., about 200 μm to about300 μm, about 300 μm to about 400 μm, about 400 μm to about 500 μm,about 500 μm to about 600 μm, about 600 μm to about 700 μm, about 700 μmto about 800 μm, about 200 μm to about 400 μm, about 200 μm to about 500μm, about 200 μm to about 600 μm, about 200 μm to about 700 μm, about200 μm to about 800 μm, about 300 μm to about 500 μm, about 300 μm toabout 600 μm, about 300 μm to about 700 μm, or about 300 μm to about 800μm.

In other aspects of this embodiment, a fluid composition comprising acrosslinked matrix polymer is sized by sieving wherein after sizing themean particle size of the crosslinked matrix polymer is, e.g., about 200μm, about 250 μm, about 300 μm, about 350 μm, about 400 μm, about 450μm, about 500 μm, about 550 μm, about 600 μm, about 650 μm, about 700μm, about 750 μm, or about 800 μm. In yet other aspects of thisembodiment, a fluid composition comprising a crosslinked matrix polymeris sized by sieving wherein after sizing the mean particle size of thecrosslinked matrix polymer is, e.g., at most 200 μm, at most 250 μm, atmost 300 μm, at most 350 μm, at most 400 μm, at most 450 μm, at most 500μm, at most 550 μm, at most 600 μm, at most 650 μm, at most 700 μm, atmost 750 μm, or at most 800 μm. In still other aspects of thisembodiment, a fluid composition comprising a crosslinked matrix polymeris sized by sieving wherein after sizing the mean particle size of thecrosslinked matrix polymer is, e.g., about 200 μm to about 300 μm, about300 μm to about 400 μm, about 400 μm to about 500 μm, about 500 μm toabout 600 μm, about 600 μm to about 700 μm, about 700 μm to about 800μm, about 200 μm to about 400 μm, about 200 μm to about 500 μm, about200 μm to about 600 μm, about 200 μm to about 700 μm, about 200 μm toabout 800 μm, about 300 μm to about 500 μm, about 300 μm to about 600μm, about 300 μm to about 700 μm, or about 300 μm to about 800 μm.

In other aspects of this embodiment, a fluid composition is sized byrecirculating between a first vessel and a second vessel through anarrow aperture wherein after recirculation the mean particle size ofthe crosslinked matrix polymer is, e.g., about 200 μm, about 250 μm,about 300 μm, about 350 μm, about 400 μm, about 450 μm, about 500 μm,about 550 μm, about 600 μm, about 650 μm, about 700 μm, about 750 μm, orabout 800 μm. In yet other aspects of this embodiment, a fluidcomposition is sized by recirculating between a first vessel and asecond vessel through a narrow aperture once wherein after recirculationthe mean particle size of the crosslinked matrix polymer is, e.g., atmost 200 μm, at most 250 μm, at most 300 μm, at most 350 μm, at most 400μm, at most 450 μm, at most 500 μm, at most 550 μm, at most 600 μm, atmost 650 μm, at most 700 μm, at most 750 μm, or at most 800 μm. In stillother aspects of this embodiment, a fluid composition is sized byrecirculating between a first vessel and a second vessel through anarrow aperture wherein after recirculation the mean particle size ofthe crosslinked matrix polymer is, e.g., about 200 μm to about 300 μm,about 300 μm to about 400 μm, about 400 μm to about 500 μm, about 500 μmto about 600 μm, about 600 μm to about 700 μm, about 700 μm to about 800μm, about 200 μm to about 400 μm, about 200 μm to about 500 μm, about200 μm to about 600 μm, about 200 μm to about 700 μm, about 200 μm toabout 800 μm, about 300 μm to about 500 μm, about 300 μm to about 600μm, about 300 μm to about 700 μm, or about 300 μm to about 800 μm.

In other aspects of this embodiment, a fluid composition comprising acrosslinked matrix polymer is sized by recirculating between a firstvessel and a second vessel through a narrow aperture wherein afterrecirculation the mean particle size of the crosslinked matrix polymeris, e.g., about 200 μm, about 250 μm, about 300 μm, about 350 μm, about400 μm, about 450 μm, about 500 μm, about 550 μm, about 600 μm, about650 μm, about 700 μm, about 750 μm, or about 800 μm. In yet otheraspects of this embodiment, a fluid composition comprising a crosslinkedmatrix polymer is sized by recirculating between a first vessel and asecond vessel through a narrow aperture wherein after recirculation themean particle size of the crosslinked matrix polymer is, e.g., at most200 μm, at most 250 μm, at most 300 μm, at most 350 μm, at most 400 μm,at most 450 μm, at most 500 μm, at most 550 μm, at most 600 μm, at most650 μm, at most 700 μm, at most 750 μm, or at most 800 μm. In stillother aspects of this embodiment, a fluid composition comprising acrosslinked matrix polymer is sized by recirculating between a firstvessel and a second vessel through a narrow aperture wherein afterrecirculation the mean particle size of the crosslinked matrix polymeris, e.g., about 200 μm to about 300 μm, about 300 μm to about 400 μm,about 400 μm to about 500 μm, about 500 μm to about 600 μm, about 600 μmto about 700 μm, about 700 μm to about 800 μm, about 200 μm to about 400μm, about 200 μm to about 500 μm, about 200 μm to about 600 μm, about200 μm to about 700 μm, about 200 μm to about 800 μm, about 300 μm toabout 500 μm, about 300 μm to about 600 μm, about 300 μm to about 700μm, or about 300 μm to about 800 μm.

In aspects of this embodiment, a fluid composition is sized byrecirculating between a first vessel and a second vessel through anarrow aperture once. In aspects of this embodiment, a fluid compositionis sized recirculating between a first vessel and a second vesselthrough a narrow aperture, e.g., twice, three times, four times, fivetimes, six times, seven times, eight times, nine times, or ten times. Inaspects of this embodiment, a fluid composition is sized byrecirculating between a first vessel and a second vessel through anarrow aperture, e.g., twice, three times, four times, five times, sixtimes, seven times, eight times, nine times, or ten times. In otheraspects of this embodiment, a fluid composition is sized byrecirculating between a first vessel and a second vessel through anarrow aperture, e.g., twice to ten times, twice to eight times, twiceto six times, twice to four times, three times to five times, threetimes to six times, or three times to seven times.

In other aspects of this embodiment, a fluid composition is recirculatedbetween a first vessel and a second vessel through a narrow aperturehaving a diameter of, e.g., about 2 mm, about 3 mm, about 4 mm, about 5mm, about 6 mm, about 7 mm, about 8 mm, about 9 mm, or about 10 mm. Inyet other aspects of this embodiment, a fluid composition isrecirculated between a first vessel and a second vessel through a narrowaperture having a diameter of, e.g., at least 2 mm, at least 3 mm, atleast 4 mm, at least 5 mm, at least 6 mm, at least 7 mm, at least 8 mm,at least 9 mm, or at least 10 mm. In still other aspects of thisembodiment, a fluid composition is recirculated between a first vesseland a second vessel through a narrow aperture having a diameter of,e.g., at most 2 mm, at most 3 mm, at most 4 mm, at most 5 mm, at most 6mm, at most 7 mm, at most 8 mm, at most 9 mm, or at most 10 mm. Infurther aspects of this embodiment, a fluid composition is recirculatedbetween a first vessel and a second vessel through a narrow aperturehaving a diameter of, e.g., about 2 mm to about 4 mm, about 2 mm toabout 6 mm, about 2 mm to about 8 mm, or about 2 mm to about 10 mm.

Additional method steps in accordance with making a fluid compositiondisclosed in the present specification may, or may not, includedegassing the composition, filling syringes with the composition, andsterilizing the composition.

Aspects of the present specification provide, in part, a method havingan optional step of degassing a fluid composition disclosed in thepresent specification. Degassing a fluid composition disclosed in thepresent specification can be accomplished using a standard device basedon conventional techniques and may be done under vacuum. Degassing isperformed for a time period sufficient to remove the desired amount ofgases from a fluid composition. Non-liming examples of sufficient timeperiod for degassing include about 2 hours to about 8 hours.

Aspects of the present specification provide, in part, a method havingan optional step of filling a syringe with a fluid composition disclosedin the present specification. Syringes useful according to the presentdescription include any syringe known in the art for administering atherapeutically effective amount a fluid composition disclosed in thepresent specification into a dermal region, such as, e.g., a syringehaving an internal volume of about 0.4 mL to about 3.0 mL. In additionthe type of needle used in conjunction with the syringe is a needlesufficient to effectively administer a therapeutically effective amountof a fluid composition disclosed in the present specification into adermal region, such as, e.g., a diameter of between about 18 G and about40 G and a needle length of at about 2 mm or more in length.

Thus, in an embodiment, a fluid composition is filled into a syringe. Inaspects of this embodiment, a fluid composition is filled into a syringehaving an internal volume of about 0.4 mL, about 0.5 mL, about 0.6 mL,about 0.7 mL, about 0.8 mL, about 0.9 mL, about 1 mL, about 1 mL, about1.5 mL, about 2 mL, about 2.5 mL, or about 3 mL. In other aspects ofthis embodiment, a fluid composition is filled into a syringe having aninternal volume of at least 0.4 mL, at least 0.5 mL, at least 0.6 mL, atleast 0.7 mL, at least 0.8 mL, at least 0.9 mL, at least 1 mL, at least1 mL, at least 1.5 mL, at least 2 mL, at least 2.5 mL, or at least 3 mL.In yet other aspects of this embodiment, a fluid composition is filledinto a syringe having an internal volume of about 0.4 mL to about 3 mL,about 0.5 mL to about 1.5 mL, or about 0.8 mL to about 2.5 mL. Inanother aspect, a syringe filled with a fluid composition is combinedwith a needle. In aspects of this embodiment, a syringe filled with afluid composition is combined with a needle having a diameter of about18 G, about 22 G, about 25 G, about 28 G, about 30 G, about 33 G, orabout 40 G. In other aspects of this embodiment, a syringe filled with afluid composition is combined with a needle having a diameter of about18 G or smaller, about 22 G or smaller, about 25 G or smaller, about 28G or smaller, about 30 G or smaller, about 33 G or smaller, or about 40G or smaller. In yet other aspects of this embodiment, a syringe filledwith a fluid composition is combined with a needle having a diameter ofabout 18 G to about 40 G, about 22 G to about 33 G, or about 26 G toabout 40 G.

Aspects of the present specification provide, in part, a method havingan optional step of sterilizing a syringe filled with a fluidcomposition disclosed in the present specification. As used herein, theterm “sterilizing” refers to any method known in the art to effectivelykill or eliminate transmissible agents without substantially altering ofdegrading a fluid composition disclosed in the specification. Asterilized fluid composition can remain stable for about 3 months toabout 3 years. One method of sterilization of a filled syringe is byautoclave. Autoclaving can be accomplished by applying a mixture ofheat, pressure and moisture to a sample in need of sterilization. Manydifferent sterilization temperatures, pressures and cycle times can beused for this step. For example, the filled syringes may be sterilizedat a temperature of at least about 120° C. to about 130° C. or greater.Moisture may or may not be utilized. The pressure applied is in someembodiments depending on the temperature used in the sterilizationprocess. The sterilization cycle may be at least about 1 minute to about20 minutes or more.

Another method of sterilization incorporates the use of a gaseousspecies which is known to kill or eliminate transmissible agents.Preferably, ethylene oxide is used as the sterilization gas and is knownin the art to be useful in sterilizing medical devices and products.

A further method of sterilization incorporates the use of an irradiationsource which is known in the art to kill or eliminate transmissibleagents. A beam of irradiation is targeted at the syringe containing theHA composition, and the wavelength of energy kills or eliminates theunwanted transmissible agents. Preferable energy useful include, but isnot limited to ultraviolet (UV) light, gamma irradiation, visible light,microwaves, or any other wavelength or band of wavelengths which killsor eliminates the unwanted transmissible agents, preferably withoutsubstantially altering of degrading the HA composition.

Thus, in an embodiment, a syringe filled with a fluid composition issterilized. In aspects of this embodiment, a syringe filled with a fluidcomposition is sterilized by autoclaving, gas sterilization, orirradiation. In other aspects of this embodiment, a syringe filled witha fluid composition can remain stable after sterilization for about 3months, about 6 months, about 9 months, about 12 months, about 18months, about 24 months, about 30 months, or about 36 months. In yetother aspects of this embodiment, a syringe filled with a fluidcomposition can remain stable after sterilization for at least 3 months,at least 6 months, at least 9 months, at least 12 months, at least 18months, at least 24 months, at least 30 months, or at least 36 months.In still other aspects of this embodiment, a syringe filled with a fluidcomposition can remain stable after sterilization for about 3 months toabout 12 months, about 3 months to about 24 months, about 3 months toabout 36 months, about 6 months to about 12 months, about 6 months toabout 24 months, or about 6 months to about 36 months.

Aspects of the present specification provide, in part, a method ofimproving a condition of skin in an individual in need thereof, themethod comprising the steps of administering a fluid compositiondisclosed in the present specification into a dermal region of theindividual, wherein the administration improves the condition.

Aspects of the present invention provide, in part, a condition of skin.Non-limiting examples of a skin condition include dehydration, lack ofskin elasticity, roughness, lack of skin tautness, skin stretch lineand/or marks, skin paleness, skin wrinkles, and the like.

Aspects of the present invention provide, in part, improving a skincondition. Non-limiting examples of improving a skin condition includerehydrating the skin, providing increased elasticity to the skin,reducing skin roughness, making the skin tauter, reducing or eliminatingstretch lines or marks, giving the skin better tone, shine, brightnessand/or radiance to reduce paleness, reducing or eliminating wrinkles inthe skin, providing wrinkle resistance to the skin, and the like.

Thus, in an embodiment, a method of treating a skin condition comprisesthe step of administering to an individual suffering from a skincondition a fluid composition comprising a matrix polymer and astabilizing component, wherein the administration of the compositionimproves the skin condition, thereby treating the skin condition. In anaspect of this embodiment, a method of treating skin dehydrationcomprises the step of administering to an individual suffering from skindehydration a fluid composition comprising a matrix polymer and astabilizing component, wherein the administration of the compositionrehydrates the skin, thereby treating skin dehydration. In anotheraspect of this embodiment, a method of treating a lack of skinelasticity comprises the step of administering to an individualsuffering from a lack of skin elasticity a fluid composition comprisinga matrix polymer and a stabilizing component, wherein the administrationof the composition increases the elasticity of the skin, therebytreating a lack of skin elasticity. In yet another aspect of thisembodiment, a method of treating skin roughness comprises the step ofadministering to an individual suffering from skin roughness a fluidcomposition comprising a matrix polymer and a stabilizing component,wherein the administration of the composition decreases skin roughness,thereby treating skin roughness. In still another aspect of thisembodiment, a method of treating a lack of skin tautness comprises thestep of administering to an individual suffering from a lack of skintautness a fluid composition comprising a matrix polymer and astabilizing component, wherein the administration of the compositionmakes the skin tauter, thereby treating a lack of skin tautness.

In a further aspect of this embodiment, a method of treating a skinstretch line or mark comprises the step of administering to anindividual suffering from a skin stretch line or mark a fluidcomposition comprising a matrix polymer and a stabilizing component,wherein the administration of the composition reduces or eliminates theskin stretch line or mark, thereby treating a skin stretch line or mark.In another aspect of this embodiment, a method of treating skin palenesscomprises the step of administering to an individual suffering from skinpaleness a fluid composition comprising a matrix polymer and astabilizing component, wherein the administration of the compositionincreases skin tone or radiance, thereby treating skin paleness. Inanother aspect of this embodiment, a method of treating skin wrinklescomprises the step of administering to an individual suffering from skinwrinkles a fluid composition comprising a matrix polymer and astabilizing component, wherein the administration of the compositionreduces or eliminates skin wrinkles, thereby treating skin wrinkles. Inyet another aspect of this embodiment, a method of treating skinwrinkles comprises the step of administering to an individual a fluidcomposition comprising a matrix polymer and a stabilizing component,wherein the administration of the composition makes the skin resistantto skin wrinkles, thereby treating skin wrinkles.

Aspects of the present invention provide, in part, a dermal region. Asused herein, the term “dermal region” refers to the region of skincomprising the epidermal-dermal junction and the dermis including thesuperficial dermis (papillary region) and the deep dermis (reticularregion). The skin is composed of three primary layers: the epidermis,which provides waterproofing and serves as a barrier to infection; thedermis, which serves as a location for the appendages of skin; and thehypodermis (subcutaneous adipose layer). The epidermis contains no bloodvessels, and is nourished by diffusion from the dermis. The main type ofcells which make up the epidermis are keratinocytes, melanocytes,Langerhans cells and Merkels cells.

The dermis is the layer of skin beneath the epidermis that consists ofconnective tissue and cushions the body from stress and strain. Thedermis is tightly connected to the epidermis by a basement membrane. Italso harbors many Mechanoreceptor/nerve endings that provide the senseof touch and heat. It contains the hair follicles, sweat glands,sebaceous glands, apocrine glands, lymphatic vessels and blood vessels.The blood vessels in the dermis provide nourishment and waste removalfrom its own cells as well as from the Stratum basale of the epidermis.The dermis is structurally divided into two areas: a superficial areaadjacent to the epidermis, called the papillary region, and a deepthicker area known as the reticular region.

The papillary region is composed of loose areolar connective tissue. Itis named for its fingerlike projections called papillae that extendtoward the epidermis. The papillae provide the dermis with a “bumpy”surface that interdigitates with the epidermis, strengthening theconnection between the two layers of skin. The reticular region liesdeep in the papillary region and is usually much thicker. It is composedof dense irregular connective tissue, and receives its name from thedense concentration of collagenous, elastic, and reticular fibers thatweave throughout it. These protein fibers give the dermis its propertiesof strength, extensibility, and elasticity. Also located within thereticular region are the roots of the hair, sebaceous glands, sweatglands, receptors, nails, and blood vessels. Tattoo ink is held in thedermis. Stretch marks from pregnancy are also located in the dermis.

The hypodermis is not part of the skin, and lies below the dermis. Itspurpose is to attach the skin to underlying bone and muscle as well assupplying it with blood vessels and nerves. It consists of looseconnective tissue and elastin. The main cell types are fibroblasts,macrophages and adipocytes (the hypodermis contains 50% of body fat).Fat serves as padding and insulation for the body.

Aspects of the present invention provide, in part, an individual. Asused herein, the term “individual” refers to any mammal including ahuman being.

Aspects of the present invention provide, in part, administering a fluidcomposition disclosed in the present specification. As used herein, theterm “administering” means any delivery mechanism that provides a fluidcomposition comprising a matrix polymer and a stabilizing component toan individual that potentially results in a clinically, therapeutically,or experimentally beneficial result. The actual delivery mechanism usedto administer a fluid composition disclosed in the present specificationto an individual can be determined by a person of ordinary skill in theart by taking into account factors, including, without limitation, thetype of skin condition, the location of the skin condition, the cause ofthe skin condition, the severity of the skin condition, the degree ofrelief desired, the duration of relief desired, the particular fluidcomposition used, the rate of excretion of the particular fluidcomposition used, the pharmacodynamics of the particular fluidcomposition used, the nature of the other compounds included in theparticular fluid composition used, the particular route ofadministration, the particular characteristics, history and risk factorsof the individual, such as, e.g., age, weight, general health and thelike, or any combination thereof.

Thus, in an embodiment, a fluid composition comprising a matrix polymerand a stabilizing component is administered to a skin region of anindividual. In an aspect of this embodiment, a fluid compositioncomprising a matrix polymer and a stabilizing component is administeredto a skin region of an individual by injection. In another aspect ofthis embodiment, a fluid composition comprising a matrix polymer and astabilizing component is administered to a skin region of an individualby injection into a dermal region. In aspects of this embodiment, afluid composition comprising a matrix polymer and a stabilizingcomponent is administered to a skin region of an individual by injectioninto, e.g., an epidermal-dermal junction region, a papillary region, areticular region, or any combination thereof.

EXAMPLES

The following examples illustrate representative embodiments nowcontemplated, but should not be construed to limit the disclosed fluidcompositions, methods of forming such fluid compositions, and methods oftreating skin conditions using such fluid compositions.

Example 1 A Method of Making a Fluid Composition

This example illustrates how to make a fluid composition disclosed inthe present specification.

To combine a stabilizing component with a physiologically-acceptablebuffer to make a stabilizing component-buffered solution, 0.9 g ofmannitol (Cooper Pharmaceutical, Inc.) was added to 1 L of a phosphatebuffer solution (comprising 8.5 g NaCl, 0.563 g NaH₂PO₄, 0.045 gNa₂HPO₄, and water) in a 10 liter bottle, and mixed for 15 minutes. Thisstep produces a stabilizing component-buffered solution comprising aphosphate buffer solution (pH 7.2) containing 0.9 mg/mL of mannitol.

To filter the stabilizing component-buffered solution, themannitol-phosphate buffer solution made above was filtered on line witha 0.2 μm filter under pressure.

To combining a matrix polymer with the stabilizing component-bufferedsolution to hydrate the matrix polymer, about 60% of themannitol-phosphate buffer solution was poured in a new bottle anduncrosslinked sodium hyaluronan polymer having a molecular weightbetween about 2.5 MDa and about 3.0 MDa was added to themannitol-phosphate buffer solution. This was completed with theremaining 40% of the mannitol-phosphate buffer solution, giving a finalconcentration of uncrosslinked hyaluronan of about 13.5 mg/L. Thiscomposition was then gently, mechanically mixed at a speed of about 100rpm to about 200 rpm for about 8 hours to about 12 hours at a coolambient temperature of about 2° C. to about 8° C. The composition wasthen subjected to agitation by manually shaking the bottle containingthe composition for about 1 minute, followed by a period of rest forabout 30 minutes. This mixing step was repeated three additional timesbefore the composition was then allowed to rest about 8 to about 12hours.

To size a fluid composition, the fluid composition above wasrecirculated four times from one bottle into a second bottle thorough anarrow orifice having a diameter of about 3 mm and about 5 mm.

To degas a fluid composition, the composition described above wasdegassed for about four hours using standard device based onconventional techniques.

Example 2A Stabilizing Component Reduces or Prevents Degradation ofMatrix Polymer

This example illustrates that the addition of a stabilizing componentreduces or prevents degradation of a matrix polymer included in a fluidcomposition disclosed in the present specification.

Fluid compositions made for testing were prepared as described inExample 1, except that 1) 1 mL of each fluid composition was made andthe amount of mannitol added to the test fluid compositions was 0% (w/v)for control, 0.5% (w/v) and 5% (w/v).

Degradation of hyaluronan polymer included in the fluid compositions andrheological processing of the results were performed according toconventional methods. Resistance of hyaluronan polymer to degradationwas assessed in fluid composition with and without mannitol and thesecompositions were compared by analyzing the time it took for thecomposition to reach a dynamic viscosity of 5 Pa·s [T(η*=5 Pa·s)]. Thedegradation test was repeated twice and a mean dynamic viscosity [T(η*=5Pa·s)_(mean)] was calculated from the 2 tests (Table 4). In general, theshorter the time to reach a dynamic viscosity of 5 Pa·s [T(η*=5 Pa·s)],the lower the resistance of hyaluronan polymer to degradation. Thus, toshow that addition of mannitol to a fluid composition had a beneficialeffect on resistance of hyaluronan polymer to breakdown, it was shownthat T(η*=5 Pa·s)_(HA with mannitol)>T(η*=5 Pa·s)_(HA without mannitol).For each fluid composition the results of the rheological test (valuesof G′, G″, tan δ, η*=f(time)+mathematical model of curve η*=f(time))were determined and the curves obtained FIGS. 1A and 1B.

TABLE 4 Breakdown Test Mean T(η* = T(η* = Std. CV Fluid Composition Test5 Pa · s) 5 Pa · s) deviation (%) Hyaluronan (13.5 mg/L) 1 747 1099 49845 2 1452 Hyaluronan (13.5 mg/L) 1 3351 3127 317 10 5% mannitol 2 2903Hyaluronan (13.5 mg/L) 1 2933 2355 818 35 0.5% mannitol 2 1776

As shown in Table 4, hyaluronan polymer was more resistant todegradation when mannitol was included in the fluid composition ascompared to the resistance hyaluronan polymer in composition withoutmannitol (Table 4, FIGS. 1A and 1B). In fact, the higher the mannitolconcentration in a fluid composition, the longer the time required forthe hyaluronan polymer to degrade. As such, incorporating a stabilizingcomponent like mannitol into a fluid composition allowed uncrosslinkedhyaluronan polymer remained intact longer because the stabilizingcomponent increased the resistance of hyaluronan polymer to degradationFIGS. 1A and 1B. Thus, the addition of a stabilizing component to afluid composition provided protection to a matrix polymer todegradation.

Example 2B Effects of Stabilizing Component on Various Properties ofFluid Composition

This example illustrates the effects of a stabilizing component onvarious properties of a fluid composition disclosed in the presentspecification including fluid appearance, pH, osmolarity, and dynamicviscosity.

Fluid compositions made for testing were prepared as described inExample 1, except that 1) 1 mL of each fluid composition was made andthe amount of mannitol added to the test fluid compositions was 0% (w/v)for control, 0.5% (w/v), 5% (w/v), 9% (w/v), and 17% (w/v).

The appearance of the fluid compositions was visually examined. Testcompositions comprising 0% (w/v), 0.5% (w/v), 5% (w/v), and 9% (w/v)mannitol all appeared colorless and transparent, while the fluidcomposition comprising 17% (w/v) mannitol appeared white and cloudy(Table 5). These results indicate that mannitol was soluble in all fluidcompositions tested, except for the composition comprising 17% (w/v)mannitol. Furthermore, it is known that mannitol solubility in water is1 g mannitol/5.5 ml water, i.e. mass solubility of 15.4%. Takentogether, these results indicate that fluid compositions comprising amatrix polymer can include about 15.4% mannitol before this polyolbegins to precipitate out of solution.

The pH of the fluid compositions was measured using a pH meter. Testcompositions comprising 0% (w/v), 0.5% (w/v), 5% (w/v), and 9% (w/v)mannitol all had a pH of 6.9, while the fluid composition comprising 17%(w/v) had a pH of 6.8 (Table 5). These results indicate that mannitolhas no effect on the pH of a fluid composition because all pH valueswere within the measurement error range of the pH meter (±0.2).

The osmolarity of the fluid compositions was measured using anosmometer. Test compositions exhibited a wide range of osmolarity thatwas correlated with the amount of mannitol added to the composition(Table 5). In general, the osmolarity of a fluid composition increase asthe amount of mannitol increased. The results indicated that a fluidcomposition comprising a matrix polymer and 0.5% mannitol preserved therequired osmolarity for hyaluronan, namely an osmolarity range of about270 mOsm/L to about 390 mOsm/L. It general, an osmolarity range of about200 mOsm/L to about 400 mOsm/L is the range approved for viscoelasticophthalmic devices.

The data obtained from the osmolarity test were further analyzed bylinear regression. This analysis revealed that y=68.576x−17.118 and thatR²=0.9888. From this formula, it was calculated that the addition of 1%mannitol to a fluid composition comprising hyaluronan would increase theosmolarity of the composition by 52 mOsm/L to approximately 351 mOsm/L.As such, a fluid composition comprising a matrix polymer and 1% mannitolwas an effective combination that balanced the opposite effects ofimproving a fluid composition's resistance to hyaluronan degradation andthe increase in osmolarity upon addition of mannitol to the composition.

TABLE 5 Mannitol Effects on Properties of Fluid Compositions Appearanceof Fluid Osmolarity Fluid Composition Composition pH (mOsm/L) Hyaluronan(13.5 mg/L) Colorless and transparent 6.9 299 Hyaluronan (13.5 mg/L)Colorless and transparent 6.9 333 Mannitol (0.5%) Hyaluronan (13.5 mg/L)Colorless and transparent 6.9 589 Mannitol (5%) Hyaluronan (13.5 mg/L)Colorless and transparent 6.9 918 Mannitol (9%) Hyaluronan (13.5 mg/L)White and cloudy 6.8 ND Mannitol (17%)

The dynamic viscosity of the fluid compositions was measured using arheometer. Test compositions comprising 0% (w/v), 5% (w/v), and 9% (w/v)mannitol exhibited a consistent dynamic viscosity over time and allwithin 20 Pa·s of each other at 1 Pa (FIG. 2). The mean dynamicviscosity of these three fluid compositions was as follows: compositionscomprising 0% (w/v) mannitol had a mean dynamic viscosity of 162 Pa·s;compositions comprising 5% (w/v) mannitol had a mean dynamic viscosityof 152 Pa·s; compositions comprising 9% (w/v) mannitol had a meandynamic viscosity of 148 Pa·s. These results indicate that the effect ofa stabilizing component was negligible because the addition of mannitolin an amount less than or equal to about 9% only resulted in a change indynamic viscosity of about 20 Pa·s at 1 Pa.

Example 3 Effects of Sterilization on Fluid Compositions Comprising aStabilizing Component

This example illustrates the effects of sterilization on variousproperties of a fluid composition disclosed in the present specificationincluding pH, dynamic viscosity and stability of stabilizing component.

Fluid compositions made for testing were prepared as described inExample 1, except that 1) 1 mL of each fluid composition was made andthe amount of mannitol added to the test fluid compositions was 0% (w/v)and 1.1% (w/v). Fluid compositions prepared above were filled in a glasssyringe and sterilized by autoclaving at 130° C. for 3 minutes.

The pH of the fluid compositions was measured using a pH meter. Theseresults indicate that sterilization had no effect on the pH of a fluidcomposition because all pH values were within the measurement errorrange of the pH meter (±0.2) (Table 6).

The dynamic viscosity of the fluid compositions was measured before andafter sterilization using a rheometer. The difference in viscositybefore and after sterilization was 54 Pa·s for fluid compositionswithout mannitol and 61 Pa·s for fluid compositions comprising 1.1%(w/v) mannitol (Table 6). These results indicate that the addition of astabilizing component had a negligible effect on dynamic viscositybecause the difference between the two compositions was within themeasurement error range of the rheometer (±10%). As such, there was nosignificant difference in dynamic viscosity of the fluid compositionsbefore and after sterilization.

The chemical and physical stability of mannitol after sterilization wasdetermined by autoclaving 25% Mannitol (Invenex Pharmaceuticals, Itasca,Ill.) five times at 121° C. for 15 minutes. The results indicate thatmannitol was chemically and physically stable after this sterilizationregime.

TABLE 6 Effects of Sterilization on Properties of Fluid CompositionsFluid Composition Hyaluronan Hyaluronan Composition (13.5 (13.5 mg/L)Sterilization Property mg/L) Mannitol (1.1%) Before Dynamic viscosity102 Pa · s 118 Pa · s sterilization at 1 Pa pH 7.1 7.0 After Dynamicviscosity  48 Pa · s  57 Pa · s sterilization at 1 Pa pH 7.1 7.0

Example 4 Determining the Stability of Fluid Composition-Filled Syringe

This example illustrates how to determine the long-term stability of afluid composition disclosed in the present specification.

Fluid compositions were prepared as described in Example 1, except thatthe amount of mannitol added to the test fluid compositions was about 1%(w/v). The degassed fluid composition was then filled into a glasssyringe and sterilized by autoclaving at 130° C. for 3 minutes.

The resulting composition-filled syringes will be stored at about 2° C.to about 8° C. and at about 18° C. to about 22° C. to determine shelflife of the fluid compositions. The stored compositions will be testedfor various properties including dynamic viscosity, pH appearance andosmolarity at 3 months, 6 months, 9 months, 12 months, 18 months, 24months, 30 months, and 36 months. These tests will determine thestability of fluid composition-filled syringes and as such the shelflife of such products.

Example 5 Safety and Efficacy Study of Fluid Composition

This example illustrates the safety and efficacy of a fluid compositiondisclosed in the present specification.

Fluid compositions were prepared as described in Example 1, except thatthe amount of mannitol added to the test fluid compositions was about 1%(w/v). The degassed fluid composition was then filled into a glasssyringe and sterilized by autoclaving at 130° C. for 3 minutes.

The study included 39 individuals and comprised men and women betweenthe ages of 30 and 45 years. All baseline data including consent,demographics, adverse events and skin evidence was collected from eachindividual before the study began. After the baseline visit, eachindividual was subdermally injected with the fluid composition describedabove and this was considered day 0 (D0). Follow-up visits took place atday 15 (D15±7 days) for the second injection, at day 30 (D30±7 days) forthe third injection and finally at day 60 (D60±7 days) for finalassessment. Dermal administration was performed using either a depotinjection method or picotage injection method. At each study visit,individuals were examined for adverse events and skin evidence, and aquestionnaire was given to assess physician and subject satisfaction.The total study duration was 60 days.

The study evaluated efficacy of the compositions on hydration andelasticity of the skin after 60 days of use of the fluid composition ondifferent skin areas including the region around the eye, the cheekregion, the peri-oral region, and the neck region.

Skin measurements were assessed at each of the four visits from day 0 today 60, on different areas, by Skin Evidence for IOMA™ (IntuiSkin,Research Triangle Park, N.C.). Skin Evidence for IOMA is a probe-basedsystem that measures the physical and visual parameters of the skin. TheVisio Probe uses its high-resolution sensor to capture with an extremeprecision the skin images including wrinkles, sebum, hairiness, darkspots, and clogged pores/bacterial infection. The Physio Probe containshigh-technology sensors and extracts in vivo the key characteristics ofthe skin including hydration, trans-epidermal water loss (TEWL), and theskin temperature.

For each individual administered the fluid composition using a depotinjection method, the cheek, neck, peri-oral and eye regions weremeasured for hydration (%) at each visit from D0 to D60 by Skin Evidencefor IOMA™ and a mean hydration was determined at each visit (Table 7).Coefficient of anisotropy (%), arithmetical roughness (Ra), andluminance (L) were also measured for each individual at each visit fromD0 to D60 by Skin Evidence for IOMA™ and mean values calculated.Statistical analyses were performed using SAS® software version 9.1 (SASInstitute, Inc. Cary, N.C.). The results indicate that overall skinhydration of the cheek (p=0.0036), neck (p=0.0346) and peri-oral(p=0.0024) regions was significantly increased by the treatment with thefluid composition. In particular, skin hydration of the cheek region wassignificantly increased by D15 of treatment (mean of 55.8%) as comparedto D0 (mean of 50.9%) and this improvement in skin condition wasmaintain through D30 of treatment (mean of 56.4%) as compared to D0(mean of 50.9%, p=0.0262) and D60 of treatment (mean of 59.3%) ascompared to D0 (mean of 50.9%, p=0.0021). In addition, skin hydration ofthe neck region was significantly increased by D15 of treatment (mean of66.0%) as compared to D0 (mean of 59.0%) and this improvement in skincondition was maintain through D30 of treatment (mean of 66.5%) of ascompared to D0 (mean of 59.0%, p=0.0022), and D60 of treatment (mean of65.3%) as compared to D0 (mean of 59.0%, p=0.0448). Furthermore, skinhydration of the peri-oral region was significantly increased by D15 oftreatment (mean of 58.9%) as compared to D0 (mean of 52.4%) and thisimprovement in skin condition was maintain through D30 of treatment(mean of 61.2%) as compared to D0 (mean of 52.4%, p=0.0041) and D60 oftreatment (mean of 59.3%) as compared to D0 (mean of 52.4%, p=0.0467).

TABLE 7 Assessment of Fluid Composition on Hydration of Skin Regions(Depot Method) Skin Visit Region D 0 D 15 D 30 D 60 P Cheek N 23 20 1718 Mixed Mean 50.9% ± 11.0% 55.8% ± 10.5% 56.4% ± 13.9% 59.3% ± 11.4%model Median 54.3% 57.7% 55.6% 58.5% P = 0.0036 Min/Max 29.1%/68.2%36.5%/72.7% 28.0%/83.1% 41.0%/76.1% Neck N 20 19 17 16 Mixed Mean 59.0%± 9.4%  66.0% ± 9.6%  66.5% ± 11.9% 65.3% ± 9.9%  model Median 58.7%66.5% 70.0% 61.4% P = 0.0346 Min/Max 41.4%/74.7% 48.4%/81.2% 43.5%/83.2%47.4%/82.9% Peri-oral N 20 16 15 15 Mixed Mean 52.4% ± 13.5% 58.9% ±12.1% 61.2% ± 14.8% 59.3% ± 11.0% model Median 50.1% 58.4% 62.6% 61.1% P= 0.0024 Min/Max 27.4%/79.6% 37.1%/81.7% 31.0%/80.6% 37.6%/79.4% Eyes N21 16 16 14 Mixed Mean 59.6% ± 12.2% 62.9% ± 10.7% 55.8% ± 14.3% 61.4% ±10.7% model Median 56.3% 60.6% 53.7% 60.9% P = 0.2808 Min/Max42.6%/81.4% 45.4%/82.6% 19.8%/80.8% 42.4%/78.3%

For each individual administered the fluid composition using a picotageinjection method, the cheek, neck, peri-oral and eye regions weremeasured for hydration (%) at each visit from D0 to D60 by Skin Evidencefor IOMA™ and a mean hydration was determined at each visit. The resultsindicated that there was no statistically significant difference in thehydration of the cheek, neck, peri-oral, or eye regions of individualsadministered the fluid compositions using the picotage injection method.

With respect to anisotropy, the results indicate an overall improvementof skin condition from the neck and peri-oral regions when the fluidcomposition was administered using a depot injection method. Nostatistically significant difference was observed in anisotropy when thefluid compositions were administered using a picotage injection method.

With respect to skin roughness, the results indicate an overallimprovement of skin condition from the neck region. In particular, skinroughness of the neck region was significantly decreased by D30 oftreatment (median of 16.7%) as compared to D0 (median of 13.9%,p=0.0001). No statistically significant difference was observed in skinroughness when the fluid compositions were administered using a picotageinjection method.

No statistically significant differences in skin luminance were detectedfor any of the four regions examined using either the depot injectionmethod or the picotage injection method.

Treatment was well-tolerated with all adverse events related toinjection technique rather than to the product. All adverse events weretransient, with a mean duration of 4 days, with no sequels.

Regarding questionnaires designed to evaluate physicians' assessment oftreatment, physicians using the depot injection method assessed skintexture (roughness) as “improved” or “very improved” in 88.9% ofindividuals administered the fluid composition at D15, in 100% ofindividuals administered the fluid composition at D30, and in 95.7% ofindividuals administered the fluid composition at D60. Physicians usingthe depot injection method assessed skin brightness as “improved” or“very improved” in 74.1% of individuals administered the fluidcomposition at D15, in 87.5% of individuals administered the fluidcomposition at D30, and in 91.3% of individuals administered the fluidcomposition at D60. Physicians using the depot injection method assessedskin hydration as “improved” or “very improved” in 88.9% of individualsadministered the fluid composition at D15, in 100% of individualsadministered the fluid composition at D30, and in 95.7% of individualsadministered the fluid composition at D60. Physicians using the depotinjection method assessed skin appearance (color) as “improved” or “veryimproved” in 48.1% of individuals administered the fluid composition atD15, in 91.7% of individuals administered the fluid composition at D30,and in 91.3% of individuals administered the fluid composition at D60.

Regarding questionnaires designed to evaluate individuals' assessment oftreatment, individuals administered the fluid composition using thedepot injection method assessed skin texture (roughness) as “improved”or “very improved” in 72.0% of individuals administered the fluidcomposition at D15, in 94.7% of individuals administered the fluidcomposition at D30, and in 80.9% of individuals administered the fluidcomposition at D60. Individuals administered the fluid composition usingthe depot injection method assessed skin brightness as “improved” or“very improved” in 84.0% of individuals administered the fluidcomposition at D15, in 84.3% of individuals administered the fluidcomposition at D30, and in 85.7% of individuals administered the fluidcomposition at D60. Individuals administered the fluid composition usingthe depot injection method assessed skin hydration as “improved” or“very improved” in 84.0% of individuals administered the fluidcomposition at D15, in 94.8% of individuals administered the fluidcomposition at D30, and in 95.0% of individuals administered the fluidcomposition at D60. Individuals administered the fluid composition usingthe depot injection method assessed skin appearance (color) as“improved” or “very improved” in 36.0% of individuals administered thefluid composition at D15, in 42.1% of individuals administered the fluidcomposition at D30, and in 42.9% of individuals administered the fluidcomposition at D60.

Regarding questionnaires designed to evaluate individuals' assessment ofinjection method, 100% of individuals administered the fluid compositionusing the depot injection method assessed global aesthetic effect at day60 of treatment as “improved” or “very improved”, whereas only 14.3% ofindividuals administered the fluid composition using the picotageinjection method assessed global aesthetic effect at day 60 of treatmentas “improved” or “very improved.” Likewise, 100% of individualsadministered the fluid composition using the depot injection methodassessed revitalization of skin at day 60 of treatment as “improved” or“very improved”, whereas only 14.3% of individuals administered thefluid composition using the picotage injection method assessedrevitalization of skin at day 60 of treatment as “improved” or “veryimproved.” Similarly, 78.9% of individuals administered the fluidcomposition using the depot injection method assessed face fullness atday 60 of treatment as “improved” or “very improved”, whereas none ofindividuals administered the fluid composition using the picotageinjection method assessed face fullness as “improved” or “veryimproved.” All told, 95% of individuals administered the fluidcomposition using the depot injection method were delighted with thetreatment at day 60, whereas only 14.3% of individuals administered thefluid composition using the picotage injection method were delightedwith the treatment at day 60.

Overall, the fluid compositions disclosed in the present specificationhave significant efficacy on skin hydration on the cheek, neck, andperi-oral regions; significant efficacy on skin anisotropy on the neckand peri-oral regions; and significant efficacy on skin roughness on theneck region. In addition evaluation of questionnaires indicate physiciansatisfaction of aesthetic results at each visit showed that skintexture, skin brightness, skin hydration and overall skin condition were“improved” or ‘very improved’ for greater than 80% of subjects at Day60. Similarly, subject satisfaction indicated that skin roughness, skinbrightness, skin hydration and number of fine wrinkles were “improved”or ‘very improved’ for greater than 80% of subjects.

In closing, it is to be understood that although aspects of the presentspecification have been described with reference to the variousembodiments, one skilled in the art will readily appreciate that thespecific examples disclosed are only illustrative of the principles ofthe subject matter disclosed in the present specification. Therefore, itshould be understood that the disclosed subject matter is in no waylimited to a particular methodology, protocol, and/or reagent, etc.,described herein. As such, various modifications or changes to oralternative configurations of the disclosed subject matter can be madein accordance with the teachings herein without departing from thespirit of the present specification. Lastly, the terminology used hereinis for the purpose of describing particular embodiments only, and is notintended to limit the scope of the present invention, which is definedsolely by the claims. Accordingly, the present invention is not limitedto that precisely as shown and described.

Certain embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention. Ofcourse, variations on these described embodiments will become apparentto those of ordinary skill in the art upon reading the foregoingdescription. The inventor expects skilled artisans to employ suchvariations as appropriate, and the inventors intend for the invention tobe practiced otherwise than specifically described herein. Accordingly,this invention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

Groupings of alternative elements or embodiments of the inventiondisclosed herein are not to be construed as limitations. Each groupmember may be referred to and claimed individually or in any combinationwith other members of the group or other elements found herein. It isanticipated that one or more members of a group may be included in, ordeleted from, a group for reasons of convenience and/or patentability.When any such inclusion or deletion occurs, the specification is deemedto contain the group as modified thus fulfilling the written descriptionof all Markush groups used in the appended claims.

Unless otherwise indicated, all numbers expressing quantities ofingredients, properties such as molecular weight, reaction conditions,and so forth used in the specification and claims are to be understoodas being modified in all instances by the term “about.” As used herein,the term “about” means that the item, parameter or term so qualifiedencompasses a range of plus or minus ten percent above and below thevalue of the stated item, parameter or term. Accordingly, unlessindicated to the contrary, the numerical parameters set forth in thespecification and attached claims are approximations that may varydepending upon the desired properties sought to be obtained by thepresent invention. At the very least, and not as an attempt to limit theapplication of the doctrine of equivalents to the scope of the claims,each numerical parameter should at least be construed in light of thenumber of reported significant digits and by applying ordinary roundingtechniques. Notwithstanding that the numerical ranges and parameterssetting forth the broad scope of the invention are approximations, thenumerical values set forth in the specific examples are reported asprecisely as possible. Any numerical value, however, inherently containscertain errors necessarily resulting from the standard deviation foundin their respective testing measurements.

The terms “a,” “an,” “the” and similar referents used in the context ofdescribing the invention (especially in the context of the followingclaims) are to be construed to cover both the singular and the plural,unless otherwise indicated herein or clearly contradicted by context.Recitation of ranges of values herein is merely intended to serve as ashorthand method of referring individually to each separate valuefalling within the range. Unless otherwise indicated herein, eachindividual value is incorporated into the specification as if it wereindividually recited herein. All methods described herein can beperformed in any suitable order unless otherwise indicated herein orotherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g., “such as”) provided herein isintended merely to better illuminate the invention and does not pose alimitation on the scope of the invention otherwise claimed. No languagein the specification should be construed as indicating any non-claimedelement essential to the practice of the invention.

Specific embodiments disclosed herein may be further limited in theclaims using consisting of or consisting essentially of language. Whenused in the claims, whether as filed or added per amendment, thetransition term “consisting of” excludes any element, step, oringredient not specified in the claims. The transition term “consistingessentially of” limits the scope of a claim to the specified materialsor steps and those that do not materially affect the basic and novelcharacteristic(s). Embodiments of the invention so claimed areinherently or expressly described and enabled herein.

All patents, patent publications, and other publications referenced andidentified in the present specification are individually and expresslyincorporated herein by reference in their entirety for the purpose ofdescribing and disclosing, for example, the methodologies described insuch publications that might be used in connection with the presentinvention. These publications are provided solely for their disclosureprior to the filing date of the present application. Nothing in thisregard should be construed as an admission that the inventors are notentitled to antedate such disclosure by virtue of prior invention or forany other reason. All statements as to the date or representation as tothe contents of these documents is based on the information available tothe applicants and does not constitute any admission as to thecorrectness of the dates or contents of these documents.

The invention claimed is:
 1. A method of improving a skin condition ofan individual, the method comprising the steps of: administering adermal filler into a dermal region of the individual, wherein the dermalfiller comprises a hyaluronan polymer composition consisting essentiallyof uncrosslinked hyaluronic acid and mannitol, wherein the mannitol ispresent in at about 0.5% (w/v) to about 5% (w/v) of the composition,wherein the uncrosslinked hyaluronic acid contains at least 95% byweight of a hyaluronic acid having a mean molecular weight of at leastabout 2,500,000 Da, and wherein the administration improves the skincondition.
 2. The method of claim 1, wherein the skin conditions treatedis skin dehydration, a lack of skin elasticity, skin roughness, a lackof skin tautness, a skin stretch line or mark, skin paleness, or skinwrinkles.
 3. The method of claim 1 wherein the dermal filler has adynamic viscosity of about 50 Pa·s to about 150 Pa·s.
 4. The method ofclaim 1 wherein the dermal filler has an osmolarity of about 200 mOsm/Lto about 400 mOsm/L.